S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from ...S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from the leaf and the first strand of cDNA was synthesized with reverse transcription. S-adenosylmethionine synthetase gene （SAMS gene） was amplified by PCR with the first strand cDNA as template and a pair of primers which was based on constructed ESTs sequence. Full-length SAMS gene sequence was obtained by BLAST comparison. According to the analysis, completed sequence of SAMS gene was integrality. The sequence of the SAMS gene was 1 185 bp in length with an opening reading frame （ORF） encoding 394 amino acids. The cDNA sequence showed a significant homology to the SAM genes from Phaseolus lunatus （89%）, Medicago sativa （85%）. A prokaryotic expression vectors based on pET-32b had been constructed and prokaryotic expression was analyzed in order to lay a strong foundation for resist adversity function analysis through situation of genic expression analysis.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
The purpose of this study is to find a new gene resource for the researches of molecular breeding of Rosaceae plants disease-resistance. Pyrus ussuriensis Maxim is used as a starting material to clone the full-length ...The purpose of this study is to find a new gene resource for the researches of molecular breeding of Rosaceae plants disease-resistance. Pyrus ussuriensis Maxim is used as a starting material to clone the full-length cDNA of NPR1（nonexpressor of pathogenesis- related genes 1） which is a key regulator in SA （salicylic acid）-mediated systemic acquired resistance （SAR） by homologous cloning and RACE techniques. The length of the cDNA sequence was 1 767 bp, the ORF was 1 761 bp, it coded 586 amino acids, pi=5.58, the relative molecular weight was 65.009 ku, contained 19 kinds of amino acids, and had full BTB/POZ and ANK domains. Compared the homology of NPR1 gene in GenBank database, the homology with Pyrus pyrifolia, Arabidopsis thaliana, Nicotiana tabacum, Lycopersicon esculentum, Oryza sativa, Helianthus annuus were 98%, 62%, 68%, 65%, 57%, 63%. The homology offunctional area were 99%, 78%, 82%, 79%, 74%, 77%. This NPR1 gene was considered as homologic gene of Pyrus ussuriensis Maxim and named PumNPR1.展开更多
We cloned and expressed bile salt hydrolase gene ofLactobacillus plantarum M1-UVS29 in Lactococcus lactis NZ9000 successfully. Gene-specific primers for amplification of L. plantarum bsh were designed by using sequenc...We cloned and expressed bile salt hydrolase gene ofLactobacillus plantarum M1-UVS29 in Lactococcus lactis NZ9000 successfully. Gene-specific primers for amplification of L. plantarum bsh were designed by using sequence which availabled from GenBank. The production of PCR amplicon was confirmed by sequencing and cloned into pMD18-T vector, and then recombined into expression vector pNZ8148 and yielding vector pNZ8148-BSH, pNZ8148-BSH was transferred into Lactococcus lactis NZ9000. Sequencing indicated that the cloned bsh fragment contained 995 nucleotides, and shared 99.3% sequence homology with bsh gene from L. plantarum MBUL10. Cloned bsh fragment was successfully transduced into NICE expression system and confirmed by PCR and restriction digest. Recombinant BSH protein was analyzed by SDS-PAGE. The molecular weight of BSH protein was approximately 37 ku. Activity of the expressed protein was 0.77 μmol· min^-1. The successfully expressed proteins by genetic engineering technology made the function of lactic acid bacteria be abundant and laid the foundation for further researches into cholesterol-lowering lactic acid bacterium food and probiotics.展开更多
Chinese isolate of transmissible gastroenteritis virus(TGEV)was propagated and harvested in swine testicle(ST)cells.Two pairs of primers were designed according to the published sequence with Oligo 4.1 and DNasis soft...Chinese isolate of transmissible gastroenteritis virus(TGEV)was propagated and harvested in swine testicle(ST)cells.Two pairs of primers were designed according to the published sequence with Oligo 4.1 and DNasis softwares.The products of RT-PCR were named Sa and Sb,of 2.3kb and 2.1kb respectively.Sa was inserted in EcoR I and Kpn I sites after Sb was cloned in Kpn I and Pst I sites of the same pUC18 plasmid.The recombinant designated pUC-S was verified and analyzed by corresponding restriction endonuclease(RE)and nested PCR on the basis of genetic sites of S gene and physical map of pUC18 plasmid,which was identified as S gene from Chinese isolate of TGEV.展开更多
Trehalose synthase is an important functional enzyme in the synthesis of trehalose in organisms and also participates in plant stress-resistant physiological processes.The transcriptomic study showed that a trehalose-...Trehalose synthase is an important functional enzyme in the synthesis of trehalose in organisms and also participates in plant stress-resistant physiological processes.The transcriptomic study showed that a trehalose-6-phosphate synthase gene was responsive to salt and alkaline stresses in Glycine soja.To dissect the molecular mechanisms of this enzyme in plant responses to stresses,the PCR technique was used to clone a trehalose-6-phosphate synthase gene from Glycine soja and it was designated as the GsTPS9.The full-length cDNA of this gene was 2583bp which encoded 861 amino acids.The sequence and structure analyses indicated that the GsTPS9 had high homology with Glycine max GmTPS9.The qRT-PCR analysis revealed that the GsTPS9 gene was expressed in Glycine soja roots,stems and leaves,and the highest expression level was in roots;the GsTPS9 gene had different responses under the stresses of NaCl,NaHCO_(3),PEG6000,ABA,MeJA and SA.This study laid the foundation for revealing the mechanism of the TPS in plant signal transduction pathways.展开更多
The outer membrane protein, ompA, ofAeromonas veronii has a role in the virulence of the organism and is a potential candidate for vaccine development. In this study, ompA I ofAeromonas veronii strain WA106 was cloned...The outer membrane protein, ompA, ofAeromonas veronii has a role in the virulence of the organism and is a potential candidate for vaccine development. In this study, ompA I ofAeromonas veronii strain WA106 was cloned and sequenced, then, it was expressed in Escherichia coli BL21. The nucleotide sequence of ompA I gene was 1 023 base pairs （GenBank Accession NO.KC748024）, which showed 100% homology with that of A. veronii （NO.AB290200.1）. This predicted protein was composed of 340 amino acid residues. Its molecular weight was 35.78 ku and isoelectric point was 5.18. The protein was a hydrophilic protein containing alpha helix and random coil with percentage of 35.0% and 49.7%, respectively. The tertiary structure, quaternary structure prediction showed that ompA I protein contained two peptide chains. SDS-PAGE showed that the actual value of the fusion protein was consistent with the expected result. It will facilitate further study of the role of ompA I protein.展开更多
Primers and probes were established according to the sequences of the alpha-amylase genes of Bacillus. halodurans C-125, Therrnus sp. IM6501, B. stearothermophilus ET-1, and B, acidopullulytics. Primers were designed ...Primers and probes were established according to the sequences of the alpha-amylase genes of Bacillus. halodurans C-125, Therrnus sp. IM6501, B. stearothermophilus ET-1, and B, acidopullulytics. Primers were designed and a 0.2 kb DNA fragment was amplified, the fragment was successfully used for the detection of the amylase Ⅱ gene in a 2 842 bp region from Bacillus halodurans strain 38C1-1.展开更多
This study has examined the structure and function of the biliverdin reductase A (BLVRA) gene and its correlation with the green eggshell character in Jinyun ducks. The previously unknown sequences of 3' and 5'...This study has examined the structure and function of the biliverdin reductase A (BLVRA) gene and its correlation with the green eggshell character in Jinyun ducks. The previously unknown sequences of 3' and 5' ends of duck BLVRA mRNA were cloned, using RT-PCR and 5' RACE methods with four pairs of primers designed from the known coding sequence (CDS) of the duck BLVRA gene. Gene expression levels of BLVRA in the shell gland were quantified by real-time PCR, and eggshell colors were determined objectively by the reflection coefficient. The cDNA sequence of BLVRA was comprised of 1071 nucleotides, encoding a protein of 303 amino acids with an isoelectric point of 7.15 and molecular weight of 34.3 kDa. The 5' and 3' ends of the duck BLVRA gene sequence published in the GenBank were extended by 238 and 199 bp, respectively. The deduced amino acid sequence of duck BLVRA had 95.3% identity with Gallus gallus, 95.0% with Taeniopygia guttata, 70.0% with Xenopus tropicalis,61.1% with Bos taurus, 60.5% with Homo sapiens and 59.5% with Mus musculus, which indicated that the BLVRA gene was evolutionarily conserved. The relative gene expression level of BLVRA in shell glands of Jinyun ducks with white eggshells was 3 times higher than that in ducks with dark green eggshells (P<0.01). There was a significant positive correlation between reflection coefficient of eggshells and the relative expression of the BLVRA gene (r=0.719, P<0.05). Together, these findings suggest that the BLVRA gene could be a candidate gene for the molecule breeding of ducks with green eggshells.展开更多
At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software too...At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.展开更多
Nine strains resistant to five fluoroquinolones (Ciprofloxacin, Ofloxacin, Enrofloxacin, Danofloxacin, Sarafloxacin) were isolated from clinical samples and extracted the chromosomal DNA of these strains. Designed pri...Nine strains resistant to five fluoroquinolones (Ciprofloxacin, Ofloxacin, Enrofloxacin, Danofloxacin, Sarafloxacin) were isolated from clinical samples and extracted the chromosomal DNA of these strains. Designed primers to amplify the Quinolone-resistance-determining region(QRDR) of gyrB gene, then the PCR products were cloned and the sequence was analyzed. In comparison with the standarded strain NCTC5776, no mutation was found in the QRDR of gyrB gene of all resistant strains. The result indicated that the QRDR of gyrB has little relationship with fluoroquinolone resistance to salmonella.展开更多
Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtaine...Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.展开更多
The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 12...The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 1296bp open reading frame and encoded 431 amino acids. According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis, Mlo gene shared over 85% nucleotide homology and 98% amino acid homology. Finally, through semi-quantitative-PCR and fluorescence quantitative analysis, we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.展开更多
Specific primers for the MC1R gene of alpacas(GenBank EU1358800) were designed to amplify the cDNA sequence using RT-PCR to seek variation in the sequence and explore the relationship between the expression level of M...Specific primers for the MC1R gene of alpacas(GenBank EU1358800) were designed to amplify the cDNA sequence using RT-PCR to seek variation in the sequence and explore the relationship between the expression level of MC1R gene and alpaca coat color.The MC1R gene from white alpaca was cloned successfully and sequence analysis verified that the MC1R gene,encoding 317 amino acids,was 1081 bp in length.Compared with the existing sequence in GenBank,sequence identity was 99.9%and 7 mutations were found.Primers,designed from the sequence obtained,were used to assess the relative expression of MC1R in alpacas of different coat color using QRT-PCR and SPSS 13.0 software.Relative expression of MC1R in the skin of brown alpacas was 4.32 times higher than that in white alpacas after normalization with GAPDH(P【0.01),indicating that MC1R expression may be related to coat color of alpacas.展开更多
The objective of the present study was to investigate the developmental expression patterns of Insulin-like growth factor-binding protein 1 (IGFBP-1) gene in different tissues of postnatal Nanjiang Mongolian Gazelles....The objective of the present study was to investigate the developmental expression patterns of Insulin-like growth factor-binding protein 1 (IGFBP-1) gene in different tissues of postnatal Nanjiang Mongolian Gazelles. Samples of heart, liver, spleen, lung, longissimus dorsi, semimembranosus, m. triceps brachii and biceps muscle of thigh were collected from a total of 36 Nanjiang Mongolian Gazelles at the age of 0, 15, 30, 60, 90 and 120 days after birth (3 males and 3 females at each age). The CDS was sequenced and ontogeny of mRNA levels of IGFBP-1 were measured by real-time fluorescence quantitative RT-PCR. The size of IGFBP-1 ORF was 792 bp encoding 263 amino acid residues, and displayed higher nucleotide/amino acid sequence identities with other ruminants compared to non-ruminants. The levels of IGFBP-1 mRNA in liver were highest (P<0.01), levels were medium in lung, spleen and heart, and the lowest in the muscles; there were no significant differences among the muscles (P>0.05). Three expression patterns of IGFBP-1 mRNA during postnatal growth from birth to day 60 were found: consistently decreasing (liver), fluctuating as increasing then decreasing (heart) or as decreasing then increasing then decreasing (spleen, lung and muscles). The results indicate that the IGFBP-1 gene is highly conserved among species, and liver has the highest expression. It was concluded that IGFBP-1 plays important roles in early postnatal growth and is expressed in a developmental-tissue-dependent manner.展开更多
As a major raw material for the textile industry and the most important fiber crop in the world,cotton is of great significance in Chinese economy.The development of cotton fiber can be divided
By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the...By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the published P8-2 strain,the homology of the necleotide sequence is 99.92%,and that of the deduced amino acid sequence is 100%.The results indicated that gD of BHV-1 was highly conservative.展开更多
Plants have developed a complicated defense mechanism during evolution to resist the harmful pathogens they encountered.The mechanism involves the interaction of the plant resistance(R)
Verticillium dahliae Kleb.is a necrotrophic plant pathogen which causes serious soil borne vascular disease in cotton.The molecular basis the defense response of cotton to this pathogen is
基金National Science Foundation (30570990)Heilongjiang Province Educational Committee Science Research Foundation (11521023)
文摘S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from the leaf and the first strand of cDNA was synthesized with reverse transcription. S-adenosylmethionine synthetase gene （SAMS gene） was amplified by PCR with the first strand cDNA as template and a pair of primers which was based on constructed ESTs sequence. Full-length SAMS gene sequence was obtained by BLAST comparison. According to the analysis, completed sequence of SAMS gene was integrality. The sequence of the SAMS gene was 1 185 bp in length with an opening reading frame （ORF） encoding 394 amino acids. The cDNA sequence showed a significant homology to the SAM genes from Phaseolus lunatus （89%）, Medicago sativa （85%）. A prokaryotic expression vectors based on pET-32b had been constructed and prokaryotic expression was analyzed in order to lay a strong foundation for resist adversity function analysis through situation of genic expression analysis.
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.
文摘The purpose of this study is to find a new gene resource for the researches of molecular breeding of Rosaceae plants disease-resistance. Pyrus ussuriensis Maxim is used as a starting material to clone the full-length cDNA of NPR1（nonexpressor of pathogenesis- related genes 1） which is a key regulator in SA （salicylic acid）-mediated systemic acquired resistance （SAR） by homologous cloning and RACE techniques. The length of the cDNA sequence was 1 767 bp, the ORF was 1 761 bp, it coded 586 amino acids, pi=5.58, the relative molecular weight was 65.009 ku, contained 19 kinds of amino acids, and had full BTB/POZ and ANK domains. Compared the homology of NPR1 gene in GenBank database, the homology with Pyrus pyrifolia, Arabidopsis thaliana, Nicotiana tabacum, Lycopersicon esculentum, Oryza sativa, Helianthus annuus were 98%, 62%, 68%, 65%, 57%, 63%. The homology offunctional area were 99%, 78%, 82%, 79%, 74%, 77%. This NPR1 gene was considered as homologic gene of Pyrus ussuriensis Maxim and named PumNPR1.
基金Supported by the National Natural Science Fund Project(31171657)Heilongjiang Province Natural Fund Project(ZD201207)Heilongjiang Province Postdoctoral Special Funds(LBH-Q13133)
文摘We cloned and expressed bile salt hydrolase gene ofLactobacillus plantarum M1-UVS29 in Lactococcus lactis NZ9000 successfully. Gene-specific primers for amplification of L. plantarum bsh were designed by using sequence which availabled from GenBank. The production of PCR amplicon was confirmed by sequencing and cloned into pMD18-T vector, and then recombined into expression vector pNZ8148 and yielding vector pNZ8148-BSH, pNZ8148-BSH was transferred into Lactococcus lactis NZ9000. Sequencing indicated that the cloned bsh fragment contained 995 nucleotides, and shared 99.3% sequence homology with bsh gene from L. plantarum MBUL10. Cloned bsh fragment was successfully transduced into NICE expression system and confirmed by PCR and restriction digest. Recombinant BSH protein was analyzed by SDS-PAGE. The molecular weight of BSH protein was approximately 37 ku. Activity of the expressed protein was 0.77 μmol· min^-1. The successfully expressed proteins by genetic engineering technology made the function of lactic acid bacteria be abundant and laid the foundation for further researches into cholesterol-lowering lactic acid bacterium food and probiotics.
文摘Chinese isolate of transmissible gastroenteritis virus(TGEV)was propagated and harvested in swine testicle(ST)cells.Two pairs of primers were designed according to the published sequence with Oligo 4.1 and DNasis softwares.The products of RT-PCR were named Sa and Sb,of 2.3kb and 2.1kb respectively.Sa was inserted in EcoR I and Kpn I sites after Sb was cloned in Kpn I and Pst I sites of the same pUC18 plasmid.The recombinant designated pUC-S was verified and analyzed by corresponding restriction endonuclease(RE)and nested PCR on the basis of genetic sites of S gene and physical map of pUC18 plasmid,which was identified as S gene from Chinese isolate of TGEV.
基金Supported by the National Natural Science Foundation of China(31670272)Heilongjiang Provincial Natural Science Foundation(C2017014)。
文摘Trehalose synthase is an important functional enzyme in the synthesis of trehalose in organisms and also participates in plant stress-resistant physiological processes.The transcriptomic study showed that a trehalose-6-phosphate synthase gene was responsive to salt and alkaline stresses in Glycine soja.To dissect the molecular mechanisms of this enzyme in plant responses to stresses,the PCR technique was used to clone a trehalose-6-phosphate synthase gene from Glycine soja and it was designated as the GsTPS9.The full-length cDNA of this gene was 2583bp which encoded 861 amino acids.The sequence and structure analyses indicated that the GsTPS9 had high homology with Glycine max GmTPS9.The qRT-PCR analysis revealed that the GsTPS9 gene was expressed in Glycine soja roots,stems and leaves,and the highest expression level was in roots;the GsTPS9 gene had different responses under the stresses of NaCl,NaHCO_(3),PEG6000,ABA,MeJA and SA.This study laid the foundation for revealing the mechanism of the TPS in plant signal transduction pathways.
基金Supported by the Science&Technology Department of Sichuan Province(2013FZ0014)the Construction Project of Postgraduate Academic Degree in Southwest University for Nationalities(2015XWD-S071007)
文摘The outer membrane protein, ompA, ofAeromonas veronii has a role in the virulence of the organism and is a potential candidate for vaccine development. In this study, ompA I ofAeromonas veronii strain WA106 was cloned and sequenced, then, it was expressed in Escherichia coli BL21. The nucleotide sequence of ompA I gene was 1 023 base pairs （GenBank Accession NO.KC748024）, which showed 100% homology with that of A. veronii （NO.AB290200.1）. This predicted protein was composed of 340 amino acid residues. Its molecular weight was 35.78 ku and isoelectric point was 5.18. The protein was a hydrophilic protein containing alpha helix and random coil with percentage of 35.0% and 49.7%, respectively. The tertiary structure, quaternary structure prediction showed that ompA I protein contained two peptide chains. SDS-PAGE showed that the actual value of the fusion protein was consistent with the expected result. It will facilitate further study of the role of ompA I protein.
基金Natural Science Foundation of Heilongjiang Province (C9912)
文摘Primers and probes were established according to the sequences of the alpha-amylase genes of Bacillus. halodurans C-125, Therrnus sp. IM6501, B. stearothermophilus ET-1, and B, acidopullulytics. Primers were designed and a 0.2 kb DNA fragment was amplified, the fragment was successfully used for the detection of the amylase Ⅱ gene in a 2 842 bp region from Bacillus halodurans strain 38C1-1.
文摘This study has examined the structure and function of the biliverdin reductase A (BLVRA) gene and its correlation with the green eggshell character in Jinyun ducks. The previously unknown sequences of 3' and 5' ends of duck BLVRA mRNA were cloned, using RT-PCR and 5' RACE methods with four pairs of primers designed from the known coding sequence (CDS) of the duck BLVRA gene. Gene expression levels of BLVRA in the shell gland were quantified by real-time PCR, and eggshell colors were determined objectively by the reflection coefficient. The cDNA sequence of BLVRA was comprised of 1071 nucleotides, encoding a protein of 303 amino acids with an isoelectric point of 7.15 and molecular weight of 34.3 kDa. The 5' and 3' ends of the duck BLVRA gene sequence published in the GenBank were extended by 238 and 199 bp, respectively. The deduced amino acid sequence of duck BLVRA had 95.3% identity with Gallus gallus, 95.0% with Taeniopygia guttata, 70.0% with Xenopus tropicalis,61.1% with Bos taurus, 60.5% with Homo sapiens and 59.5% with Mus musculus, which indicated that the BLVRA gene was evolutionarily conserved. The relative gene expression level of BLVRA in shell glands of Jinyun ducks with white eggshells was 3 times higher than that in ducks with dark green eggshells (P<0.01). There was a significant positive correlation between reflection coefficient of eggshells and the relative expression of the BLVRA gene (r=0.719, P<0.05). Together, these findings suggest that the BLVRA gene could be a candidate gene for the molecule breeding of ducks with green eggshells.
基金Supported by the National Natural Science Foundation of China(32272724)the National Science Foundation of Heilongjiang Province,China(LH2019C033)。
文摘At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.
文摘Nine strains resistant to five fluoroquinolones (Ciprofloxacin, Ofloxacin, Enrofloxacin, Danofloxacin, Sarafloxacin) were isolated from clinical samples and extracted the chromosomal DNA of these strains. Designed primers to amplify the Quinolone-resistance-determining region(QRDR) of gyrB gene, then the PCR products were cloned and the sequence was analyzed. In comparison with the standarded strain NCTC5776, no mutation was found in the QRDR of gyrB gene of all resistant strains. The result indicated that the QRDR of gyrB has little relationship with fluoroquinolone resistance to salmonella.
基金Supported by the Postdoctoral Scientific Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.
基金Supported by Postdoctoral Scientifi c Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 1296bp open reading frame and encoded 431 amino acids. According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis, Mlo gene shared over 85% nucleotide homology and 98% amino acid homology. Finally, through semi-quantitative-PCR and fluorescence quantitative analysis, we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.
基金supported by the National Natural Science Foundation of China(No.30501070)Shanxi Natural Science Foundation(No.20041099)President Foundation of Agricultural University of Hebei (BS2007023)
文摘Specific primers for the MC1R gene of alpacas(GenBank EU1358800) were designed to amplify the cDNA sequence using RT-PCR to seek variation in the sequence and explore the relationship between the expression level of MC1R gene and alpaca coat color.The MC1R gene from white alpaca was cloned successfully and sequence analysis verified that the MC1R gene,encoding 317 amino acids,was 1081 bp in length.Compared with the existing sequence in GenBank,sequence identity was 99.9%and 7 mutations were found.Primers,designed from the sequence obtained,were used to assess the relative expression of MC1R in alpacas of different coat color using QRT-PCR and SPSS 13.0 software.Relative expression of MC1R in the skin of brown alpacas was 4.32 times higher than that in white alpacas after normalization with GAPDH(P【0.01),indicating that MC1R expression may be related to coat color of alpacas.
基金supported by Scientific Research Fund of Sichuan Province Education Department(No.09ZA073)Sichuan Provincial Technology Support Project(No.2011NZ0003)
文摘The objective of the present study was to investigate the developmental expression patterns of Insulin-like growth factor-binding protein 1 (IGFBP-1) gene in different tissues of postnatal Nanjiang Mongolian Gazelles. Samples of heart, liver, spleen, lung, longissimus dorsi, semimembranosus, m. triceps brachii and biceps muscle of thigh were collected from a total of 36 Nanjiang Mongolian Gazelles at the age of 0, 15, 30, 60, 90 and 120 days after birth (3 males and 3 females at each age). The CDS was sequenced and ontogeny of mRNA levels of IGFBP-1 were measured by real-time fluorescence quantitative RT-PCR. The size of IGFBP-1 ORF was 792 bp encoding 263 amino acid residues, and displayed higher nucleotide/amino acid sequence identities with other ruminants compared to non-ruminants. The levels of IGFBP-1 mRNA in liver were highest (P<0.01), levels were medium in lung, spleen and heart, and the lowest in the muscles; there were no significant differences among the muscles (P>0.05). Three expression patterns of IGFBP-1 mRNA during postnatal growth from birth to day 60 were found: consistently decreasing (liver), fluctuating as increasing then decreasing (heart) or as decreasing then increasing then decreasing (spleen, lung and muscles). The results indicate that the IGFBP-1 gene is highly conserved among species, and liver has the highest expression. It was concluded that IGFBP-1 plays important roles in early postnatal growth and is expressed in a developmental-tissue-dependent manner.
基金This work was funded by grants fromthe National Basic Research and Development Program(2004CB117304)the Hi-tech Research and Development Program of China (2007AA10Z115)
文摘As a major raw material for the textile industry and the most important fiber crop in the world,cotton is of great significance in Chinese economy.The development of cotton fiber can be divided
文摘By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the published P8-2 strain,the homology of the necleotide sequence is 99.92%,and that of the deduced amino acid sequence is 100%.The results indicated that gD of BHV-1 was highly conservative.
文摘Plants have developed a complicated defense mechanism during evolution to resist the harmful pathogens they encountered.The mechanism involves the interaction of the plant resistance(R)
文摘Verticillium dahliae Kleb.is a necrotrophic plant pathogen which causes serious soil borne vascular disease in cotton.The molecular basis the defense response of cotton to this pathogen is
