Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expres...Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro- MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly （P〈0.05）. After knocked down MSTN gene, P21 expression decreased （P〈0.05）, but CDK2 gene expression increased （P〈0.05）. These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.展开更多
For solving the dilemma of the short exothermic life-span of WO_(3)based metastable interstitial composites(MICs)with extensive application prospect,this paper has firstly designed the promising antiwetting Al/WO_(3)M...For solving the dilemma of the short exothermic life-span of WO_(3)based metastable interstitial composites(MICs)with extensive application prospect,this paper has firstly designed the promising antiwetting Al/WO_(3)MICs via electrophoresis assembly of nano-Al and WO_(3)particles fabricated by hydrothermal synthesis method,followed by the subsequent fluorination treatment.A combination of X ray diffraction(XRD),field emission scanning electron microscope(FESEM),energy dispersive X-ray spectroscopy(EDX),and Fourier transform infrared spectroscopy(FT-IR)techniques were utilized in order to characterize the crystal structure,microstructure,and elemental composition distribution of target films after different natural exposure tests.The product with uniform distribution and high purity possesses a high contact angle of~170°and a minute sliding angle of~1°,and displays the outstanding anti-wetting property using droplets with different surface tensions.It also shows great moisture stability in high relative-humidity circumstances after one year of the natural exposure experiment.Notably,the heat output of a fresh sample can reach up to 2.3 kJ/g and retain 96%after the whole exposure test,showing outstanding thermo-stability for at least one year.This work further proposed the mechanism of antiwetting Al/WO_(3)MICs considering the variation tendency of their DSC curve,providing a valuable theoretical reference for designing other self-protected MICs with a long exothermic life-span applied in wide fields of national defense,military industry,etc.展开更多
The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this ...The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this research field obtained over the past two years.This report summarizes the major progress of the basic researches about the origin of life in China during 2018–2020.展开更多
The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose⁃catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monit...The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose⁃catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16℃in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni⁃NTA affinity column.Size⁃exclusion chromatography and SDS⁃PAGE analysis demonstrated that the puri⁃fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD^(+)as a coenzyme to NADP^(+).The optimal temperature and pH of the TeXDH were 40℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26%of the initial enzyme activity,while the divalent metal ions Mg^(2+)or Ca^(2+)could recover the enzyme activity of TeXDH,reaching 103.32%and 110.69%of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn^(2+),Ni^(2+),Cu^(2+),Zn^(2+),Co^(2+),and Cd^(2+)significantly inhibited the activity of TeXDH.ICP⁃MS and molecular doc⁃king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn^(2+)ions and 1 mol/L Mg^(2+)ion.Further⁃more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.展开更多
4-Nonylphenol(NP)is a kind of estrogen belonging to the endocrine disrupter,widely used in various agricultural and industrial goods.However,extensive use of NP with direct release to environment poses high risks to b...4-Nonylphenol(NP)is a kind of estrogen belonging to the endocrine disrupter,widely used in various agricultural and industrial goods.However,extensive use of NP with direct release to environment poses high risks to both human health and ecosystems.Herein,for the first time,we developed near-infrared(NIR)responsive upconversion luminescence nanosensor for NP detection.The Förster resonance energy transfer based upconversion nanoparticles(UCNPs)-graphene oxide sensor offers highly selective and sensitive detection of NP in linear ranges of 5−200 ng/mL and 200−1000 ng/mL under 980 nm and 808 nm excitation,respectively,with LOD at 4.2 ng/mL.The sensors were successfully tested for NP detection in real liquid milk samples with excellent recovery results.The rare-earth fluoride based upconversion luminescence nanosensor with NIR excitation wavelength,holds promise for sensing food,environmental,and biological samples due to their high sensitivity,specific recognition,low LOD,negligible autofluorescence,along with the deep penetration of NIR excitation sources.展开更多
Nicotine,also known as nicotinic norephedrine,is one of the main alkaloids present in tobacco plants.In recent years,due to the increase in tobacco production and smoking population,the environmental and health issues...Nicotine,also known as nicotinic norephedrine,is one of the main alkaloids present in tobacco plants.In recent years,due to the increase in tobacco production and smoking population,the environmental and health issues caused by nicotine have become increasingly severe.Traditional methods have proven ineffective in efficiently degrading residual nicotine.To address this issue,scientists both domestically and internationally have turned to biodegradation methods to tackle the environmental and health problems caused by residual nicotine.In this study,an enrichment method was used to screen bacteria with nicotine-degrading capabilities from the soil of tobacco planting sites at the Tobacco Research Institute of Heilongjiang in Bin County,Harbin City.Through phenotypic observations and 16S rDNA identification,a bacterial strain identified as Pseudomonas hunanensis MGJ-2 was isolated,capable of utilizing nicotine as a carbon and nitrogen source for growth.High-performance liquid chromatography(HPLC)-1 analysis revealed that within 25 h,strain MGJ-2 could degrade nicotine 500 mg·L^(-1) with an efficiency exceeding 99.9%.Strain MGJ-2 was applied to tobacco,and after 15 days of incubation and fermentation,it degraded 10.57%of nicotine in tobacco.Overall,the discovery of strain MGJ-2 enriched the resources of nicotine-degrading strains.Its remarkable biodegradation performance held immense potential for future biodegradation of nicotine in tobacco.展开更多
In order to elucidate the relationship between the pathogen carriage rate in seeds of muskmelon(Cucumis melo L.)and the incidence rate of Fusarium wilt of muskmelon(FWM),as well as to identify potential biological con...In order to elucidate the relationship between the pathogen carriage rate in seeds of muskmelon(Cucumis melo L.)and the incidence rate of Fusarium wilt of muskmelon(FWM),as well as to identify potential biological control agents against FWM,this study conducted both pot and field experiments to evaluate the efficacy of Gongzhulingmycin on FWM and its impact on muskmelon yield.The results indicated that the pathogen carriage rates of different species in muskmelon seeds varied significantly,showing a positive correlation with disease incidence during the seedling stage.The results from pot tests indicated that in comparison to the control,disease indices were significantly reduced following treatment with prochloraz and 100 times Gongzhulingmycin at both 7 days and 14 days post-emergence of FWM symptoms.Concurrently,root growth was enhanced.Field experiment outcomes demonstrated that relative to the control,there was a decrease in FWM incidence during the fruit-setting stage,along with an increase in theoretical output per square kilometer for muskmelon.Although the efficacy of Gongzhulingmycin against FWM was lower than that observed for prochloraz,it exhibited significant effects on biomass enhancement and disease resistance.Therefore,it showed promise as a potential biological control agent for managing FWM.展开更多
Platycodon grandiflorum A.DC.(PAl)C)root was taken as experiment material to extract polysaccharide.On the base of single factor tests(extraction time,extraction temperature,liquid-solid ratio,solvent pH value and NaC...Platycodon grandiflorum A.DC.(PAl)C)root was taken as experiment material to extract polysaccharide.On the base of single factor tests(extraction time,extraction temperature,liquid-solid ratio,solvent pH value and NaCl concentration),the study concluded the main factors affecting the extraction of PADC polysaccharide,which are liquid-solid ratio,extraction time and extraction temperature.Then through central-composite test design,the extraction conditions were concluded as liquid-solid ratio 34.43,extraction time 89.83 min and extraction temperature 52.47℃.By means of validation experiments,the adequacy of this model was confirmed.展开更多
Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescen...Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescence material Ru(bpy)32+·6H2O, and then the particles were characterized via transmission electron microscope. The fluorescence nanoparticles were conjugated with plasmid DNA to form complexes, and then treated with ultrasound and DNase I. pEGAD plasmid DNA-nanoparticle complexes were co-cultured with plant suspension cells of Dioscrea Zigiberensis G H Wright, and treated with ultrasound. The results show that the diameter of the fluorescence starch-nanoparticles is 50-100 nm. DNA-nanoparticle complexes can protect DNA from ultrasound damage as well as from DNase I cleavage. Mediated by ultrasound, pEGAD plasmid DNA-nanoparticle complexes can pierce into the cell wall, cell membrane and nucleus membrane of plant suspension cells. The green fluorescence protein(GFP) gene at a high frequency exceeds 5%. This nano-biomaterial can efficiently solve the problem that exterior genes cannot traverse the plant cell wall easily.展开更多
Methionine （Met） and lysine （Lys） have been reported as the first two limiting amino acids （AA） for maximum milk yield and milk protein production. Supplying these AA may improve microbial protein synthesis and ...Methionine （Met） and lysine （Lys） have been reported as the first two limiting amino acids （AA） for maximum milk yield and milk protein production. Supplying these AA may improve microbial protein synthesis and therefore improve milk production without adding excess N to the environment. This observation utilized fermented soybean meal （SBM）, cottonseed meal （CSM）, rapeseed meal （RSM） and corn by Bacillus subtilis 168 and Leuconostoc mesenteroides as core feedstuffs to produce special biological protein feed for dairy cow. The results showed that the milk production, milk protein percentage, milk fat percentage and milk DM percentage of test groups in trial period were significantly more than those of the control group （P〈0.01）, the results showed that adding fermenting protein feed in dairy cow diets could significantly improve milk yield, milk protein and milk fat content. The economic benefits of actual application were analyzed, the group of 0.5% was the best compared with the other groups.展开更多
The myeloblastosis oncogenes(MYB)are important transcription factors that facilitate induction of variously develop-mental and stress responsive genes.They are hence,emerging as key players in improving stress toleran...The myeloblastosis oncogenes(MYB)are important transcription factors that facilitate induction of variously develop-mental and stress responsive genes.They are hence,emerging as key players in improving stress tolerance of plants in response to several abiotic stresses.It was predicted that DfMYB2 contained an open reading frame(ORF)of 1621 bp coding 377 amino acid residues with molecular weight of 41.5 ku.It had 50 potential phosphorylation sites,44 potential N-glycosylation sites and one trans-membrane domain.In neighbor-joining(NJ)phylogenetic tree,DfMYB2 was close to the branch of angiosperms.The subcellular localization of DfMYB2 was in nucleus.The expressions of DfMYB2 increased gradually in the filament,prothalli and sporophyte.The results showed that DfMYB2 played a more and more important role in its growth and development.And the expression levels in rhizomes were significantly higher than those in roots and leaves.After abscisic acid(ABA)and PEG600 treatment,DfMYB2 showed a downward trend,followed by an upward trend.The expression of DfMYB2 was inhibited in a short time under drought stress,the strong induction of DfMYB2 expression by ABA indicated that it was possibly involved in abiotic stress responses in an ABA-dependent manner.展开更多
For the first time, we used Tullgren method made a study on vertical migrating and cluster analysis of the soil mesofauna in Dongying Halophytes Garden in the Yellow River Delta （YRD）, Shandong Province. The results...For the first time, we used Tullgren method made a study on vertical migrating and cluster analysis of the soil mesofauna in Dongying Halophytes Garden in the Yellow River Delta （YRD）, Shandong Province. The results showed that the soil mesofauna tended to gather on soil surface in most samples at most times, but the vertical migrating greatly varied in different seasons or environment conditions. Acari was the dominant group. The index of diversity of the soil fauna was correlated with the index of evenness. The Acari＇s number of individuals infected other species and numbers. Dominant group-Aeari made greater contribution to the result of cluster analysis, and there were significant differences between communities in different habitats by cluster analysis with both Bray-Curtis and Jaccard similarity coefficient.展开更多
Connected a segment of CMV enhancer to the front of MyoG gene promoter and then constructed the corresponding dual luciferase expression vector pGL3-CMV-MyoGpro. We set four eukaryotic expression vectors including pGL...Connected a segment of CMV enhancer to the front of MyoG gene promoter and then constructed the corresponding dual luciferase expression vector pGL3-CMV-MyoGpro. We set four eukaryotic expression vectors including pGL3-CMV, pGL3MyoGpro, pGL3-CMV-MyoGpro, and pGL3-Basic which contained CMV promoter, MyoG promoter, CMV-MyoG synthesis promoter, and a promoterless negative control, respectively. Then the four vectors and internal control Renilla luciferase report gene vector phRL-TK were transfected into bovine skeletal muscle satellite cells, mouse C2C12 cells and bovine fetal fibroblast cells to detect the promoter activity with dual luciferase report system. The results showed that CMV enhancer could significantly improve the transcription activity of bovine MyoG gene promoter in muscle satellite cells and mouse C2C12 cells, and it had certain specificity. This study provided experimental materials for increasing the high expression of exogenous gene in bovine muscle cells, and also laid the molecular theoretical basis for obtaining the high specific promoter of bovine muscle and the transgenic beef cattle.展开更多
According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gen...According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gene was gotten from Bacillus bifidus ATCC 29521 by PCR. BSH gene was inserted into lactic acid bacteria expression vector pNZ8148 to construct the recombinant pNZ8148-BSH. The recombinant pNZ8148-BSH was transferred into lactic acid bacteria NZ9000 with electrotransformation method. And the recombinant which could express BSH protein was obtained. It was identified by SDS-PAGE electrophoresis and activity verification. The result could provide a rationale reference for expressing BSH in lactic acid bacteria.展开更多
Iron(Ⅱ) tetrasulfophthalocyanine(FeTSPc) ,as a novel mimetic enzyme of peroxidase,was used in the synthesis of a conducting polyaniline(PANI) /sodium dodecylsulfate(SDS) complex in SDS aqueous micellar solutions. The...Iron(Ⅱ) tetrasulfophthalocyanine(FeTSPc) ,as a novel mimetic enzyme of peroxidase,was used in the synthesis of a conducting polyaniline(PANI) /sodium dodecylsulfate(SDS) complex in SDS aqueous micellar solutions. The effects of pH,concentrations of aniline,SDS and H2O2,and reaction time on polymerization of aniline were studied in this case as shown by UV-Vis absorption spectroscopy. The results show that a wide range of pH(0.5-4.0) is required to produce the conducting PANI,and the optimal pH is 1.0 in SDS micelle. The optimal concentrations of aniline,SDS and H2O2 in feed,and reaction time in this case for the production of conducting PANI are respectively 10 mmol/L,10 mmol/L,25 mmol/L,and 15 h. FT-IR spectrum,elemental analysis,conductivity,cyclic voltammetry and thermogravimetric analysis confirm the thermal stability and electroactive form of PANI.展开更多
Lactobacillus was selected as a bacterial carrier for expression of N-lobe of porcine lactoferrin （PLFN）. A pair of primers was designed with Oligo6.0 and used to amplify PLFN gene. It was in accordance with the cha...Lactobacillus was selected as a bacterial carrier for expression of N-lobe of porcine lactoferrin （PLFN）. A pair of primers was designed with Oligo6.0 and used to amplify PLFN gene. It was in accordance with the characters of translational fusions from gene and expression vector plasmid. A 1 077 bp fragment of the gene from PLF was cloned from mammary gland tissue of the lactating sow on the third day by RT-PCR; the gene was connected with the vector plasmid pPG612.1 and transformed into the host strain JM109. The recombinant expression vector plasmid pPG612-PLFN was created and identified by using plasmid extraction, PCR, restriction enzyme digestion and sequence analysis. The recombinant plasmid was transformed into Lactobacillus casei ATCC393, Lactobacillus plantarum KLDS 1.0344, Lactobacillus paracasei KLDS 1.0652 and Lactobacillus pentosus KLDS 1.0413 by electroporation, and produced the recombinant strains of pPG612-PLFN/L, casei, pPG612-PLFN/L, plantarum, pPG612-PLFN/ L. paracasei and pPG612-PLFN/L, pentosus, respectively. The results indicated that PLFN gene had inserted into the expression vectors and achieved multiple Laetobacillus expression systems. It electes the base for the expression and production of recombinant porcine lactoferrin in Lactobaeillus展开更多
To examine the effect of myogenin gene on the differentiation of bovine skeletal muscle satellite cell, we constructed small interfering RNA plasmid vector to obtain myogenin knockdown bovine skeletal muscle cells, th...To examine the effect of myogenin gene on the differentiation of bovine skeletal muscle satellite cell, we constructed small interfering RNA plasmid vector to obtain myogenin knockdown bovine skeletal muscle cells, then used cell transfection, real time RCR and Western Blot to detect the influence of myogenin to cell differentiation. Results showed that the knockdown of myogenin significantly decreased its expression and other muscle-specific genes. Compared to the control, it could differentiate into few myotubes when challenged by low serum in the medium. These findings provided an important theoretical basis for further explore of the genetic mechanism in adult skeletal muscle, the remedy of muscle injuries and the cultivation of high-yield transgenic cattle.展开更多
MicroRNAs （miRNAs） are one kind of small RNA in all eukaryote. MicroRNAs can regulate gene expression of eukaryote; they widely participate in every physiological process. They can block mRNA expression or cleave mR...MicroRNAs （miRNAs） are one kind of small RNA in all eukaryote. MicroRNAs can regulate gene expression of eukaryote; they widely participate in every physiological process. They can block mRNA expression or cleave mRNA by complement to target mRNA. Scholars estimate miRNA genes occuping about 1% of genome, but they can regulate 10%-30% genes of whole genome. The genes are regulated by miRNA including signal proteins, enzymes, transcription factors, and so on. In the field of plant research, the start of miRNA research is later, but it is proved that plant miRNAs are important to every plant physiological process. Now miRNA has become the hotspot of plant molecular biology research. This paper introduced the biology function, action mechanism, researching method and recently development of microRNAs, also focused on advances in plant microRNAs. This paper has important reference value for plant stress tolerance miRNA research.展开更多
Winter wheat （Triticum aestivum） cultivars Dongnongdongmai 1 with strong cold/freezing tolerance and Jimai 22 with weak cold/freezing tolerance were used for investigating the difference of microstructure and ultras...Winter wheat （Triticum aestivum） cultivars Dongnongdongmai 1 with strong cold/freezing tolerance and Jimai 22 with weak cold/freezing tolerance were used for investigating the difference of microstructure and ultrastructure between leaves of two cultivars under low temperature stress （5℃ and -15 ℃） using optical and electron microscope. The results showed that there was no obvious difference between leaves of Dongnongdongmai 1 and Jimai 22 in microstructure. However, the difference between those leaves was distinct in ultrastructure. The grana lamella and stroma lamella were stacked regularly and arranged parallelly along the long axis of chloroplasts in cv. Dongnongdongmai 1, while the arrangement directions of thylakoids in Jimai 22＇s leaves were so irregular as to form various angles with the long axis of chloroplasts. At -15℃, the mitochondrias were swelled to be round and the structure of cristaes became blurry in both cultivars＇ leaves, while some cristaes of Jimai 22 disappeared. These results would provide theoretical evidence for selecting cold/freezing tolerant winter wheat germplasm resources展开更多
基金Supported by the Major Special Projects of New Product Training of Transgenic Organisms(zx080072008-2008)
文摘Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro- MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly （P〈0.05）. After knocked down MSTN gene, P21 expression decreased （P〈0.05）, but CDK2 gene expression increased （P〈0.05）. These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.
基金funded by the financial support from National Natural Science Foundation of China(Grant No 21805014 and No82102635)Science and Technology Research Project of Chongqing Education Board(Grant No.KJQN201901428)。
文摘For solving the dilemma of the short exothermic life-span of WO_(3)based metastable interstitial composites(MICs)with extensive application prospect,this paper has firstly designed the promising antiwetting Al/WO_(3)MICs via electrophoresis assembly of nano-Al and WO_(3)particles fabricated by hydrothermal synthesis method,followed by the subsequent fluorination treatment.A combination of X ray diffraction(XRD),field emission scanning electron microscope(FESEM),energy dispersive X-ray spectroscopy(EDX),and Fourier transform infrared spectroscopy(FT-IR)techniques were utilized in order to characterize the crystal structure,microstructure,and elemental composition distribution of target films after different natural exposure tests.The product with uniform distribution and high purity possesses a high contact angle of~170°and a minute sliding angle of~1°,and displays the outstanding anti-wetting property using droplets with different surface tensions.It also shows great moisture stability in high relative-humidity circumstances after one year of the natural exposure experiment.Notably,the heat output of a fresh sample can reach up to 2.3 kJ/g and retain 96%after the whole exposure test,showing outstanding thermo-stability for at least one year.This work further proposed the mechanism of antiwetting Al/WO_(3)MICs considering the variation tendency of their DSC curve,providing a valuable theoretical reference for designing other self-protected MICs with a long exothermic life-span applied in wide fields of national defense,military industry,etc.
文摘The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this research field obtained over the past two years.This report summarizes the major progress of the basic researches about the origin of life in China during 2018–2020.
基金湖南省教育厅基金优秀青年项目(No.22B0482)湖南科技大学博士启动基金(No.E51992 and E51993)资助。
文摘The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose⁃catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16℃in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni⁃NTA affinity column.Size⁃exclusion chromatography and SDS⁃PAGE analysis demonstrated that the puri⁃fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD^(+)as a coenzyme to NADP^(+).The optimal temperature and pH of the TeXDH were 40℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26%of the initial enzyme activity,while the divalent metal ions Mg^(2+)or Ca^(2+)could recover the enzyme activity of TeXDH,reaching 103.32%and 110.69%of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn^(2+),Ni^(2+),Cu^(2+),Zn^(2+),Co^(2+),and Cd^(2+)significantly inhibited the activity of TeXDH.ICP⁃MS and molecular doc⁃king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn^(2+)ions and 1 mol/L Mg^(2+)ion.Further⁃more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.
文摘4-Nonylphenol(NP)is a kind of estrogen belonging to the endocrine disrupter,widely used in various agricultural and industrial goods.However,extensive use of NP with direct release to environment poses high risks to both human health and ecosystems.Herein,for the first time,we developed near-infrared(NIR)responsive upconversion luminescence nanosensor for NP detection.The Förster resonance energy transfer based upconversion nanoparticles(UCNPs)-graphene oxide sensor offers highly selective and sensitive detection of NP in linear ranges of 5−200 ng/mL and 200−1000 ng/mL under 980 nm and 808 nm excitation,respectively,with LOD at 4.2 ng/mL.The sensors were successfully tested for NP detection in real liquid milk samples with excellent recovery results.The rare-earth fluoride based upconversion luminescence nanosensor with NIR excitation wavelength,holds promise for sensing food,environmental,and biological samples due to their high sensitivity,specific recognition,low LOD,negligible autofluorescence,along with the deep penetration of NIR excitation sources.
基金Supported by the Research on Biofermentation Technology of Domestic Cigar Tobacco(202115010534-JS-178)(2021)。
文摘Nicotine,also known as nicotinic norephedrine,is one of the main alkaloids present in tobacco plants.In recent years,due to the increase in tobacco production and smoking population,the environmental and health issues caused by nicotine have become increasingly severe.Traditional methods have proven ineffective in efficiently degrading residual nicotine.To address this issue,scientists both domestically and internationally have turned to biodegradation methods to tackle the environmental and health problems caused by residual nicotine.In this study,an enrichment method was used to screen bacteria with nicotine-degrading capabilities from the soil of tobacco planting sites at the Tobacco Research Institute of Heilongjiang in Bin County,Harbin City.Through phenotypic observations and 16S rDNA identification,a bacterial strain identified as Pseudomonas hunanensis MGJ-2 was isolated,capable of utilizing nicotine as a carbon and nitrogen source for growth.High-performance liquid chromatography(HPLC)-1 analysis revealed that within 25 h,strain MGJ-2 could degrade nicotine 500 mg·L^(-1) with an efficiency exceeding 99.9%.Strain MGJ-2 was applied to tobacco,and after 15 days of incubation and fermentation,it degraded 10.57%of nicotine in tobacco.Overall,the discovery of strain MGJ-2 enriched the resources of nicotine-degrading strains.Its remarkable biodegradation performance held immense potential for future biodegradation of nicotine in tobacco.
基金Supported by the Key Research and Development Project of the Department of Science and Technology of Jilin Province(20230203175SF)。
文摘In order to elucidate the relationship between the pathogen carriage rate in seeds of muskmelon(Cucumis melo L.)and the incidence rate of Fusarium wilt of muskmelon(FWM),as well as to identify potential biological control agents against FWM,this study conducted both pot and field experiments to evaluate the efficacy of Gongzhulingmycin on FWM and its impact on muskmelon yield.The results indicated that the pathogen carriage rates of different species in muskmelon seeds varied significantly,showing a positive correlation with disease incidence during the seedling stage.The results from pot tests indicated that in comparison to the control,disease indices were significantly reduced following treatment with prochloraz and 100 times Gongzhulingmycin at both 7 days and 14 days post-emergence of FWM symptoms.Concurrently,root growth was enhanced.Field experiment outcomes demonstrated that relative to the control,there was a decrease in FWM incidence during the fruit-setting stage,along with an increase in theoretical output per square kilometer for muskmelon.Although the efficacy of Gongzhulingmycin against FWM was lower than that observed for prochloraz,it exhibited significant effects on biomass enhancement and disease resistance.Therefore,it showed promise as a potential biological control agent for managing FWM.
文摘Platycodon grandiflorum A.DC.(PAl)C)root was taken as experiment material to extract polysaccharide.On the base of single factor tests(extraction time,extraction temperature,liquid-solid ratio,solvent pH value and NaCl concentration),the study concluded the main factors affecting the extraction of PADC polysaccharide,which are liquid-solid ratio,extraction time and extraction temperature.Then through central-composite test design,the extraction conditions were concluded as liquid-solid ratio 34.43,extraction time 89.83 min and extraction temperature 52.47℃.By means of validation experiments,the adequacy of this model was confirmed.
基金Project(200501) supported the "985" Program of China
文摘Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescence material Ru(bpy)32+·6H2O, and then the particles were characterized via transmission electron microscope. The fluorescence nanoparticles were conjugated with plasmid DNA to form complexes, and then treated with ultrasound and DNase I. pEGAD plasmid DNA-nanoparticle complexes were co-cultured with plant suspension cells of Dioscrea Zigiberensis G H Wright, and treated with ultrasound. The results show that the diameter of the fluorescence starch-nanoparticles is 50-100 nm. DNA-nanoparticle complexes can protect DNA from ultrasound damage as well as from DNase I cleavage. Mediated by ultrasound, pEGAD plasmid DNA-nanoparticle complexes can pierce into the cell wall, cell membrane and nucleus membrane of plant suspension cells. The green fluorescence protein(GFP) gene at a high frequency exceeds 5%. This nano-biomaterial can efficiently solve the problem that exterior genes cannot traverse the plant cell wall easily.
基金Supported by"863"Project of Ministry of Science and Technology of China(2013AA102504-03)
文摘Methionine （Met） and lysine （Lys） have been reported as the first two limiting amino acids （AA） for maximum milk yield and milk protein production. Supplying these AA may improve microbial protein synthesis and therefore improve milk production without adding excess N to the environment. This observation utilized fermented soybean meal （SBM）, cottonseed meal （CSM）, rapeseed meal （RSM） and corn by Bacillus subtilis 168 and Leuconostoc mesenteroides as core feedstuffs to produce special biological protein feed for dairy cow. The results showed that the milk production, milk protein percentage, milk fat percentage and milk DM percentage of test groups in trial period were significantly more than those of the control group （P〈0.01）, the results showed that adding fermenting protein feed in dairy cow diets could significantly improve milk yield, milk protein and milk fat content. The economic benefits of actual application were analyzed, the group of 0.5% was the best compared with the other groups.
基金Supported by the National Natural Science Foundation of China(31570189)。
文摘The myeloblastosis oncogenes(MYB)are important transcription factors that facilitate induction of variously develop-mental and stress responsive genes.They are hence,emerging as key players in improving stress tolerance of plants in response to several abiotic stresses.It was predicted that DfMYB2 contained an open reading frame(ORF)of 1621 bp coding 377 amino acid residues with molecular weight of 41.5 ku.It had 50 potential phosphorylation sites,44 potential N-glycosylation sites and one trans-membrane domain.In neighbor-joining(NJ)phylogenetic tree,DfMYB2 was close to the branch of angiosperms.The subcellular localization of DfMYB2 was in nucleus.The expressions of DfMYB2 increased gradually in the filament,prothalli and sporophyte.The results showed that DfMYB2 played a more and more important role in its growth and development.And the expression levels in rhizomes were significantly higher than those in roots and leaves.After abscisic acid(ABA)and PEG600 treatment,DfMYB2 showed a downward trend,followed by an upward trend.The expression of DfMYB2 was inhibited in a short time under drought stress,the strong induction of DfMYB2 expression by ABA indicated that it was possibly involved in abiotic stress responses in an ABA-dependent manner.
基金Supported by the Doctoral Fund of Northeast Agricultural University(2009RC41)Postdoctoral Grants of Heilongjiang Province(LBH-Z10265)
文摘For the first time, we used Tullgren method made a study on vertical migrating and cluster analysis of the soil mesofauna in Dongying Halophytes Garden in the Yellow River Delta （YRD）, Shandong Province. The results showed that the soil mesofauna tended to gather on soil surface in most samples at most times, but the vertical migrating greatly varied in different seasons or environment conditions. Acari was the dominant group. The index of diversity of the soil fauna was correlated with the index of evenness. The Acari＇s number of individuals infected other species and numbers. Dominant group-Aeari made greater contribution to the result of cluster analysis, and there were significant differences between communities in different habitats by cluster analysis with both Bray-Curtis and Jaccard similarity coefficient.
基金Supported by the Major Special Projects of New Product Training of Transgenic Organisms(zx080072008-2008)
文摘Connected a segment of CMV enhancer to the front of MyoG gene promoter and then constructed the corresponding dual luciferase expression vector pGL3-CMV-MyoGpro. We set four eukaryotic expression vectors including pGL3-CMV, pGL3MyoGpro, pGL3-CMV-MyoGpro, and pGL3-Basic which contained CMV promoter, MyoG promoter, CMV-MyoG synthesis promoter, and a promoterless negative control, respectively. Then the four vectors and internal control Renilla luciferase report gene vector phRL-TK were transfected into bovine skeletal muscle satellite cells, mouse C2C12 cells and bovine fetal fibroblast cells to detect the promoter activity with dual luciferase report system. The results showed that CMV enhancer could significantly improve the transcription activity of bovine MyoG gene promoter in muscle satellite cells and mouse C2C12 cells, and it had certain specificity. This study provided experimental materials for increasing the high expression of exogenous gene in bovine muscle cells, and also laid the molecular theoretical basis for obtaining the high specific promoter of bovine muscle and the transgenic beef cattle.
基金Supported by 863 Projects (2008AA10Z311)National Science and Technology Support Projects (2009BADB9B06)+1 种基金Started Post-doctoral Research Grant of Heilongjiang Province (LBH-Q07023)Harbin Technological Innovation of Special Funds (2007RFQXN020)
文摘According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gene was gotten from Bacillus bifidus ATCC 29521 by PCR. BSH gene was inserted into lactic acid bacteria expression vector pNZ8148 to construct the recombinant pNZ8148-BSH. The recombinant pNZ8148-BSH was transferred into lactic acid bacteria NZ9000 with electrotransformation method. And the recombinant which could express BSH protein was obtained. It was identified by SDS-PAGE electrophoresis and activity verification. The result could provide a rationale reference for expressing BSH in lactic acid bacteria.
基金Supported by the National Natural Science Foundation of China（30270924） Natural Science Foundation of Anhui Province （070411004） +1 种基金 the National Science Foundation of Education Department of Anhui Province（2006KJ184B） the Foundation of Provincial Key Lab of Biotic Environment and Ecological Safety in Anhui Province
基金Project(07JJ6020) supported by the Hunan Provincial Natural Science Foundation of ChinaProject(2007-24-3) supported by the Huaihua Key Science and Technology Program, China
文摘Iron(Ⅱ) tetrasulfophthalocyanine(FeTSPc) ,as a novel mimetic enzyme of peroxidase,was used in the synthesis of a conducting polyaniline(PANI) /sodium dodecylsulfate(SDS) complex in SDS aqueous micellar solutions. The effects of pH,concentrations of aniline,SDS and H2O2,and reaction time on polymerization of aniline were studied in this case as shown by UV-Vis absorption spectroscopy. The results show that a wide range of pH(0.5-4.0) is required to produce the conducting PANI,and the optimal pH is 1.0 in SDS micelle. The optimal concentrations of aniline,SDS and H2O2 in feed,and reaction time in this case for the production of conducting PANI are respectively 10 mmol/L,10 mmol/L,25 mmol/L,and 15 h. FT-IR spectrum,elemental analysis,conductivity,cyclic voltammetry and thermogravimetric analysis confirm the thermal stability and electroactive form of PANI.
基金Supported by National Natural Science Foundation of China (30871809,31072057)Principal Fund of Northeast Agricultural University
文摘Lactobacillus was selected as a bacterial carrier for expression of N-lobe of porcine lactoferrin （PLFN）. A pair of primers was designed with Oligo6.0 and used to amplify PLFN gene. It was in accordance with the characters of translational fusions from gene and expression vector plasmid. A 1 077 bp fragment of the gene from PLF was cloned from mammary gland tissue of the lactating sow on the third day by RT-PCR; the gene was connected with the vector plasmid pPG612.1 and transformed into the host strain JM109. The recombinant expression vector plasmid pPG612-PLFN was created and identified by using plasmid extraction, PCR, restriction enzyme digestion and sequence analysis. The recombinant plasmid was transformed into Lactobacillus casei ATCC393, Lactobacillus plantarum KLDS 1.0344, Lactobacillus paracasei KLDS 1.0652 and Lactobacillus pentosus KLDS 1.0413 by electroporation, and produced the recombinant strains of pPG612-PLFN/L, casei, pPG612-PLFN/L, plantarum, pPG612-PLFN/ L. paracasei and pPG612-PLFN/L, pentosus, respectively. The results indicated that PLFN gene had inserted into the expression vectors and achieved multiple Laetobacillus expression systems. It electes the base for the expression and production of recombinant porcine lactoferrin in Lactobaeillus
基金Supported by the Ministry of Agricultural Nuarture of New Varieties Genetically Modified Organisms Significant Special Funding (2008ZX08007-002)
文摘To examine the effect of myogenin gene on the differentiation of bovine skeletal muscle satellite cell, we constructed small interfering RNA plasmid vector to obtain myogenin knockdown bovine skeletal muscle cells, then used cell transfection, real time RCR and Western Blot to detect the influence of myogenin to cell differentiation. Results showed that the knockdown of myogenin significantly decreased its expression and other muscle-specific genes. Compared to the control, it could differentiate into few myotubes when challenged by low serum in the medium. These findings provided an important theoretical basis for further explore of the genetic mechanism in adult skeletal muscle, the remedy of muscle injuries and the cultivation of high-yield transgenic cattle.
文摘MicroRNAs （miRNAs） are one kind of small RNA in all eukaryote. MicroRNAs can regulate gene expression of eukaryote; they widely participate in every physiological process. They can block mRNA expression or cleave mRNA by complement to target mRNA. Scholars estimate miRNA genes occuping about 1% of genome, but they can regulate 10%-30% genes of whole genome. The genes are regulated by miRNA including signal proteins, enzymes, transcription factors, and so on. In the field of plant research, the start of miRNA research is later, but it is proved that plant miRNAs are important to every plant physiological process. Now miRNA has become the hotspot of plant molecular biology research. This paper introduced the biology function, action mechanism, researching method and recently development of microRNAs, also focused on advances in plant microRNAs. This paper has important reference value for plant stress tolerance miRNA research.
基金Supported by Doctoral Scientific Research Project of Northeast Agricultural University (20082010)Innovative Research Team Project of Northeast Agricultural University (CXZ003)
文摘Winter wheat （Triticum aestivum） cultivars Dongnongdongmai 1 with strong cold/freezing tolerance and Jimai 22 with weak cold/freezing tolerance were used for investigating the difference of microstructure and ultrastructure between leaves of two cultivars under low temperature stress （5℃ and -15 ℃） using optical and electron microscope. The results showed that there was no obvious difference between leaves of Dongnongdongmai 1 and Jimai 22 in microstructure. However, the difference between those leaves was distinct in ultrastructure. The grana lamella and stroma lamella were stacked regularly and arranged parallelly along the long axis of chloroplasts in cv. Dongnongdongmai 1, while the arrangement directions of thylakoids in Jimai 22＇s leaves were so irregular as to form various angles with the long axis of chloroplasts. At -15℃, the mitochondrias were swelled to be round and the structure of cristaes became blurry in both cultivars＇ leaves, while some cristaes of Jimai 22 disappeared. These results would provide theoretical evidence for selecting cold/freezing tolerant winter wheat germplasm resources
