Background Polygalacturonase inhibiting proteins(PGIPs)play a pivotal role in plant defense against plant patho-gens by inhibiting polygalacturonase(PG),an enzyme produced by pathogens to degrade plant cell wall pecti...Background Polygalacturonase inhibiting proteins(PGIPs)play a pivotal role in plant defense against plant patho-gens by inhibiting polygalacturonase(PG),an enzyme produced by pathogens to degrade plant cell wall pectin.PGIPs,also known as leucine-rich repeat pathogenesis-related(PR)proteins,activate the host’s defense response upon interaction with PG,thereby reinforcing the host defense against plant pathogens attacks.In Egyptian or extra-long staple cotton(Gossypium barbadense),the interaction between PGIP and PG is one of the crucial steps in the defense mechanism against major pathogens such as Xanthomonas citri pv.malvacearum and Alternaria mac-rospora,which are responsible for bacterial leaf blight and leaf spot diseases,respectively.Results To unravel the molecular mechanisms underlying these PR proteins,we conducted a comprehensive study involving molecular modeling,protein-protein docking,site-specific double mutation(E169G and F242K),and molec-ular dynamics simulations.Both wild-type and mutated cotton PGIPs were examined in the interaction with the PG enzyme of a bacterial and fungal pathogen.Our findings revealed that changes in conformations of double-mutated residues in the active site of PGIP lead to the inhibition of PG binding.The molecular dynamics simulation studies provide insights into the dynamic behaviour and stability of the PGIP-PG complexes,shedding light on the intricate details of the inhibitory and exhibitory mechanism against the major fungal and bacterial pathogens of G.barbadense,respectively.Conclusions The findings of this study not only enhance our understanding of the molecular interactions between PGs of Xanthomonas citri pv.malvacearum and Alternaria macrospora and PGIP of G.barbadense but also pre-sent a potential strategy for developing the disease-resistant cotton varieties.By variations in the binding affinities of PGs through specific mutations in PGIP,this research offers promising avenues for the development of enhanced resistance to cotton plants against bacterial leaf blight and leaf spot diseases.展开更多
Creutzfeldt-Jakob disease(CJD)is a rare neurodegenerative disorder characterized by abnormalities in the prion protein(PrP),the most common form of human prion disease.Although Genome-Wide Association Studies(GWAS)hav...Creutzfeldt-Jakob disease(CJD)is a rare neurodegenerative disorder characterized by abnormalities in the prion protein(PrP),the most common form of human prion disease.Although Genome-Wide Association Studies(GWAS)have identified numerous risk genes for CJD,the mechanisms underlying these risk loci remain poorly understood.This study aims to elucidate novel genetically prioritized candidate proteins associated with CJD in the human brain through an integrative analytical pipeline.Utilizing datasets from Protein Quantitative Trait Loci(pQTL)(NpQTL1=152,NpQTL2=376),expression QTL(eQTL)(N=452),and the CJD GWAS(NCJD=4110,NControls=13569),we implemented a systematic analytical pipeline.This pipeline included Proteome-Wide Association Study(PWAS),Mendelian randomization(MR),Bayesian colocalization,and Transcriptome-Wide Association Study(TWAS)to identify novel genetically prioritized candidate proteins implicated in CJD pathogenesis within the brain.Through PWAS,we identified that the altered abundance of six brain proteins was significantly associated with CJD.Two genes,STX6 and PDIA4,were established as lead causal genes for CJD,supported by robust evidence(False Discovery Rate<0.05 in MR analysis;PP4/(PP3+PP4)≥0.75 in Bayesian colocalization).Specifically,elevated levels of STX6 and PDIA4 were associated with an increased risk of CJD.Additionally,TWAS demonstrated that STX6 and PDIA4 were associated with CJD at the transcriptional level.展开更多
Background Transgenic research in crops involves using genetic engineering techniques to introduce specific genes of interest from other organisms,or even entirely new genes into plant genomes to create crops with des...Background Transgenic research in crops involves using genetic engineering techniques to introduce specific genes of interest from other organisms,or even entirely new genes into plant genomes to create crops with desirable traits that wouldn’t be possible through conventional breeding methods.Transgenic crops have been developed for various traits globally.Whitefly,Bemisia tabaci(Gennadius)is one of the major sucking pests of cotton that cause significant damage to the cotton production.To combat whitefly infestations,researchers have developed four transgenic cotton lines expressing the fern protein.And those transgenic lines need to be evaluated for their performance against the target pest—whitefly.The evaluation was designed as controlled trials in polyhouse or muslin cloth cages under open-choice and no-choice conditions by comparing four transgenic cotton lines(A,B,C,and D)with three control groups,including untransformed cotton plants with a same genetic background of the transgenic line,conventionally bred whitefly-resistant cotton,and whitefly-susceptible cotton.In order to study the generational effect,the evaluation also involved studies on whitefly development in laboratory,muslin cloth cage,and polyhouse conditions.Results Both open-choice and no-choice experiments had shown that all the four transgenic cotton lines(A,B,C,and D)expressing the fern protein reduced adult whitefly numbers significantly compared with the control lines,except for the no-choice conditions in 2021,where the transgenic line C was non-significant different from the resistant control line.Notably,the nymphal population on the resistant control line was relatively low and nonsignificant different from the transgenic line C in 2021;and the transgenic lines A and C in 2022 under open-choice conditions.Under no-choice condition,the nymphal counts in the resistant control line was non-significant different from transgenic lines C and D in 2021;and transgenic line D in 2022.All transgenic lines showed significant decrease in egg hatching in 2021 and nymphal development in 2022,except for the transgenic line C which had no significant different in the nymphal development comparing with non-transgenic control lines in 2022.Adult emergence rates in both years of evaluation showed significant decrease in transgenic lines A and B comparing with the control lines.Additionally,the results showed a significant reduction in cotton leaf curl disease and sooty mold development in all the four transgenic lines compared with susceptible control under open-choice conditions,indicating potential benefits of transgenic lines beyond direct effect on whitefly control.Furthermore,the research explored the generational effects of the fern protein on whitefly which revealed the lowest fecundity in the transgenic line C across F0,F1 and F3 generations,lower egg hatching in F1 and F2 generations in transgenic lines A and B,shorter nymphal duration in F1 and F2 generations in transgenic line B,and the least total adult emergence in the transgenic line C in F0 and F3 generations.Conclusions These findings suggest that the transgenic cotton lines expressing fern protein disrupts whitefly populations and the life cycle to a certain extent.However,results are not consistent over generations and years of study,indicating these transgenic lines were not superior over control lines and need to be improved in future breeding.展开更多
The current study comprehensively evaluates four different protein extraction methods based on urea,sodium dodecyl sulfate(SDS),anionic surfactants(BT),and total RNA extractor(Trizol),aiming to optimize the sample pre...The current study comprehensively evaluates four different protein extraction methods based on urea,sodium dodecyl sulfate(SDS),anionic surfactants(BT),and total RNA extractor(Trizol),aiming to optimize the sample preparation workflow for mass spectrometry-based proteomics.Using HeLa cells as an example,we found that the method employing the mass spectrometry-compatible surfactant BT reagent significantly reduces the total time consumed for protein extraction and minimizes protein losses during the sample preparation process.Further integrating the four protein extraction methods,we identified over 7000 proteins from HeLa cells without relying on pre-fractionation techniques,and 2990 of them were quantified using label-free quantification.It is worth noting that the BT and SDS methods demonstrate higher efficiency in extracting membrane proteins,while the Urea and Trizol methods are more effective in extracting proteins from nuclear and cytoplasmic fractions.In summary,this study provides a novel solution for deep proteome coverage,particularly in the context of cellular protein extraction,by integrating mass spectrometry-compatible surfactants with traditional extraction methods to effectively enhance protein identification numbers.展开更多
Profiling the protein composition of bacteria is essential for understanding their biology,physiology and interaction with environment.Mass spectrometry has become a pivotal tool for protein analysis,facilitating the ...Profiling the protein composition of bacteria is essential for understanding their biology,physiology and interaction with environment.Mass spectrometry has become a pivotal tool for protein analysis,facilitating the examination of expression levels,molecular masses and structural modifications.In this study,we compared the performance of three widely-used mass spectrometry methods,i.e.,matrix-assisted laser desorption/ionization(MALDI)protein fingerprinting,top-down proteomics and bottom-up proteomics,in the profiling of bacterial protein composition.It was revealed that bottom-up proteomics provided the highest protein coverage and exhibited the greatest protein profile overlap between bacterial species.In contrast,MALDI protein fingerprinting demonstrated superior detection reproducibility and effectiveness in distinguishing between bacterial species.Although top-down proteomics identified fewer proteins than bottom-up approach,it complemented MALDI fingerprinting in the discovery of bacterial protein markers,both favoring abundant,stable,and hydrophilic bacterial ribosomal proteins.This study represents the most systematic and comprehensive comparison of mass spectrometry-based protein profiling methodologies to date.It provides valuable guidelines for the selection of appropriate profiling strategies for specific analytical purposes.This will facilitate studies across various fields,including infection diagnosis,antimicrobial resistance detection and pharmaceutical target discovery.展开更多
The assembly of a protein complex is very important for its biological function,which can be investigated by determining the order of assembly/disassembly of its protein subunits.Although static structures of many pro...The assembly of a protein complex is very important for its biological function,which can be investigated by determining the order of assembly/disassembly of its protein subunits.Although static structures of many protein com-plexes are available in the protein data bank,their assembly/disassembly orders of subunits are largely unknown.In addition to experimental techniques for studying subcomplexes in the assembly/disassembly of a protein complex,computational methods can be used to predict the assembly/disassembly order.Since sampling is a nontrivial issue in simulating the assembly/disassembly process,coarse-grained simulations are more efficient than atomic simulations are.In this work,we developed computational protocols for predicting the assembly/disassembly orders of protein complexes via coarse-grained simulations.The protocols were illustrated via two protein complexes,and the predicted assembly/disassembly orders were consistent with the available experimental data.展开更多
目的研究尘螨肠道微生物蛋白Hypothetical protein CE2118的表达和纯化,为研究其在尘螨疫苗免疫治疗中的作用奠定基础。方法采用生物信息学方法,根据GenBank中Hypothetical protein CE2118蛋白的基因序列,将其中稀有密码子改造为大肠杆...目的研究尘螨肠道微生物蛋白Hypothetical protein CE2118的表达和纯化,为研究其在尘螨疫苗免疫治疗中的作用奠定基础。方法采用生物信息学方法,根据GenBank中Hypothetical protein CE2118蛋白的基因序列,将其中稀有密码子改造为大肠杆菌常用密码子并进行二级结构优化,合成Hypothetical protein CE2118基因,构建原核表达载体pGEX6P-1-hypothetical protein CE2118并经酶切鉴定,在大肠埃希菌Rosetta(DE3)中用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,重组产物采用GST亲和层析纯化柱纯化。结果经密码子改造和二级结构优化后Hypothetical protein CE2118基因长度为588bp,其编码蛋白理论分子量为21kDa。重组表达载体经酶切鉴定与理论推测结果相符,该基因经IPTG诱导在大肠埃希菌Rosetta(DE3)中得到高效的可溶性表达,纯化后的重组蛋白分子量约为21kDa,其单一蛋白纯度达95%以上。结论本研究成功构建了Hypothetical protein CE2118基因的pGEX6P-1原核重组质粒,获得的可溶性重组蛋白为进一步研究肠道微生物蛋白Hypothetical protein CE2118在尘螨疫苗免疫治中的作用机理奠定基础。展开更多
In winter, spring and summer, the rhizome of wild Elytrzgia repens of Heilongjiang Province was selected to extract the soluble which whole protein and the apoplastic protein, and analyzed by SDS-PAGE. The result indi...In winter, spring and summer, the rhizome of wild Elytrzgia repens of Heilongjiang Province was selected to extract the soluble which whole protein and the apoplastic protein, and analyzed by SDS-PAGE. The result indicated that there were two specific polypeptides in two types protein from winter; their relative molecular weight were identified as 52 ku and 26 ku by analyzing software; the apoplastic protein from winter had the ability of modifing the growth of ice crystal which appeared hexagonal in shape observed with the phase-contrast photomicroscope. So the apoplastic protein from winter has the antifreeze characters and the 52 ku protein is more likely the antifreeze protein展开更多
Background Increased levels of inflammatory markers have been documented in various settings of coronary artery disease. The vulnerability of coronary lesions in acute myocardial infarction(AMI) at the time of onset m...Background Increased levels of inflammatory markers have been documented in various settings of coronary artery disease. The vulnerability of coronary lesions in acute myocardial infarction(AMI) at the time of onset may be related to serum levels of C reactive protein(CRP) on admission, before CRP levels are affected by myocardial damage.Objective This study assessed the predictive value of CRP levels within six hours after the onset of acute anterior myocardial infarction with primary percutaneous coronary intervention(PCI).Methods The plasma CRP of 76 patients with first acute anterior myocardial infarction was measured within 6 hours after onset. They were divided into 2 groups: group 1( n =20) with elevated CRP( ≥0.3mg/dl ) on admission within 6 hours after onset and group 2( n =56) with normal CRP( <0.3mg/dl ) within 6 hours after onset. All patients were treated by primary PCI. The primary combined end points, including death due to cardiac causes, re MI related to the infarction artery(RIA) and repeat intervention of the RIA, and the restenosis rate were assessed in relation to CRP levels within 6 hours after onset. Left ventricular end diastolic volume index(EDVI),end systolic volume index(ESVI),and ejection fraction(EF) on admission and 6 month after the onset were assessed by left ventriculography. Changes in EDVI(ΔEDVI),ESVI(ΔESVI), and EF(ΔEF) were obtained by subtracting respective on admission values from corresponding 6 month follow up values. Results There were no significant differences in baseline characteristics between the two groups. The primary combined end points were significantly more frequent in group 1(20%) than those in group 2( 1.79% , P <0.01 ).In addition, restenosis rates were significantly higher in group 1 than in group 2(41.18% vs 16.07%, P<0.05). Group 1 showed greater increases in left ventricular volume and less improvement in EF compared with group 2(ΔEDVI 6.31 ±2.17 vs 3.29 ±9.46ml/m 2 , ΔESVI 5.92 ±2.31 vs 3.86 ±1.08ml/m 2 , ΔEF 1.92 ±0.47 vs 4.79 ±1.73% , P <0.05 , respectively).Conclusions CRP levels within 6 hours after the onset of AMI might predict adverse outcome after primary PCI and progressive ventricular remodeling within 6 month of AMI.展开更多
We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constr...We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.展开更多
This pilot study involved the application of a crossflow ultrafiltration （UF） membrane module employing hollow fiber polysulphone membranes in the reclamation of protein and oligosaccharides from soy protein wastewa...This pilot study involved the application of a crossflow ultrafiltration （UF） membrane module employing hollow fiber polysulphone membranes in the reclamation of protein and oligosaccharides from soy protein wastewater. The optimal membrane molecular weight cut-off （MWCO） was selected as 10 ku upon retaining ratios of protein and oligosaccharides as well as the variation of permeate flux. The effects of pretreatment strategy and operating conditions, such as transmembrane pressure （TMP）, feed pH and velocity on flux were studied. According to the experimental result, the optimal operating conditions were determined as temperature 45 ℃, pH 4.5, TMP 0.2 MPa and feed velocity 10 L·s^-1. In addition, the permeate flux recovery by pure water backflushing and chemical cleaning was investigated. The experimental result showed that backflushing with pure water could only recover the lost permeate flux by 30%, and the decrease of backflushing interval was helpful in improving UF permeate productivity. Chemical cleaning test revealed that the combination of sodium hydroxide and EDTA was an ideal agent for cleaning the fouled membranes.展开更多
基金CABin grant(F.no.Agril.Edn.4-1/2013-A&P)Indian Council of Agricul-tural Research,Ministry of Agriculture and Farmers’Welfare,Govt.of India and Department of Biotechnology,Govt.of India for BIC project grant(BT/PR40161/BTIS/137/32/2021)。
文摘Background Polygalacturonase inhibiting proteins(PGIPs)play a pivotal role in plant defense against plant patho-gens by inhibiting polygalacturonase(PG),an enzyme produced by pathogens to degrade plant cell wall pectin.PGIPs,also known as leucine-rich repeat pathogenesis-related(PR)proteins,activate the host’s defense response upon interaction with PG,thereby reinforcing the host defense against plant pathogens attacks.In Egyptian or extra-long staple cotton(Gossypium barbadense),the interaction between PGIP and PG is one of the crucial steps in the defense mechanism against major pathogens such as Xanthomonas citri pv.malvacearum and Alternaria mac-rospora,which are responsible for bacterial leaf blight and leaf spot diseases,respectively.Results To unravel the molecular mechanisms underlying these PR proteins,we conducted a comprehensive study involving molecular modeling,protein-protein docking,site-specific double mutation(E169G and F242K),and molec-ular dynamics simulations.Both wild-type and mutated cotton PGIPs were examined in the interaction with the PG enzyme of a bacterial and fungal pathogen.Our findings revealed that changes in conformations of double-mutated residues in the active site of PGIP lead to the inhibition of PG binding.The molecular dynamics simulation studies provide insights into the dynamic behaviour and stability of the PGIP-PG complexes,shedding light on the intricate details of the inhibitory and exhibitory mechanism against the major fungal and bacterial pathogens of G.barbadense,respectively.Conclusions The findings of this study not only enhance our understanding of the molecular interactions between PGs of Xanthomonas citri pv.malvacearum and Alternaria macrospora and PGIP of G.barbadense but also pre-sent a potential strategy for developing the disease-resistant cotton varieties.By variations in the binding affinities of PGs through specific mutations in PGIP,this research offers promising avenues for the development of enhanced resistance to cotton plants against bacterial leaf blight and leaf spot diseases.
文摘Creutzfeldt-Jakob disease(CJD)is a rare neurodegenerative disorder characterized by abnormalities in the prion protein(PrP),the most common form of human prion disease.Although Genome-Wide Association Studies(GWAS)have identified numerous risk genes for CJD,the mechanisms underlying these risk loci remain poorly understood.This study aims to elucidate novel genetically prioritized candidate proteins associated with CJD in the human brain through an integrative analytical pipeline.Utilizing datasets from Protein Quantitative Trait Loci(pQTL)(NpQTL1=152,NpQTL2=376),expression QTL(eQTL)(N=452),and the CJD GWAS(NCJD=4110,NControls=13569),we implemented a systematic analytical pipeline.This pipeline included Proteome-Wide Association Study(PWAS),Mendelian randomization(MR),Bayesian colocalization,and Transcriptome-Wide Association Study(TWAS)to identify novel genetically prioritized candidate proteins implicated in CJD pathogenesis within the brain.Through PWAS,we identified that the altered abundance of six brain proteins was significantly associated with CJD.Two genes,STX6 and PDIA4,were established as lead causal genes for CJD,supported by robust evidence(False Discovery Rate<0.05 in MR analysis;PP4/(PP3+PP4)≥0.75 in Bayesian colocalization).Specifically,elevated levels of STX6 and PDIA4 were associated with an increased risk of CJD.Additionally,TWAS demonstrated that STX6 and PDIA4 were associated with CJD at the transcriptional level.
文摘Background Transgenic research in crops involves using genetic engineering techniques to introduce specific genes of interest from other organisms,or even entirely new genes into plant genomes to create crops with desirable traits that wouldn’t be possible through conventional breeding methods.Transgenic crops have been developed for various traits globally.Whitefly,Bemisia tabaci(Gennadius)is one of the major sucking pests of cotton that cause significant damage to the cotton production.To combat whitefly infestations,researchers have developed four transgenic cotton lines expressing the fern protein.And those transgenic lines need to be evaluated for their performance against the target pest—whitefly.The evaluation was designed as controlled trials in polyhouse or muslin cloth cages under open-choice and no-choice conditions by comparing four transgenic cotton lines(A,B,C,and D)with three control groups,including untransformed cotton plants with a same genetic background of the transgenic line,conventionally bred whitefly-resistant cotton,and whitefly-susceptible cotton.In order to study the generational effect,the evaluation also involved studies on whitefly development in laboratory,muslin cloth cage,and polyhouse conditions.Results Both open-choice and no-choice experiments had shown that all the four transgenic cotton lines(A,B,C,and D)expressing the fern protein reduced adult whitefly numbers significantly compared with the control lines,except for the no-choice conditions in 2021,where the transgenic line C was non-significant different from the resistant control line.Notably,the nymphal population on the resistant control line was relatively low and nonsignificant different from the transgenic line C in 2021;and the transgenic lines A and C in 2022 under open-choice conditions.Under no-choice condition,the nymphal counts in the resistant control line was non-significant different from transgenic lines C and D in 2021;and transgenic line D in 2022.All transgenic lines showed significant decrease in egg hatching in 2021 and nymphal development in 2022,except for the transgenic line C which had no significant different in the nymphal development comparing with non-transgenic control lines in 2022.Adult emergence rates in both years of evaluation showed significant decrease in transgenic lines A and B comparing with the control lines.Additionally,the results showed a significant reduction in cotton leaf curl disease and sooty mold development in all the four transgenic lines compared with susceptible control under open-choice conditions,indicating potential benefits of transgenic lines beyond direct effect on whitefly control.Furthermore,the research explored the generational effects of the fern protein on whitefly which revealed the lowest fecundity in the transgenic line C across F0,F1 and F3 generations,lower egg hatching in F1 and F2 generations in transgenic lines A and B,shorter nymphal duration in F1 and F2 generations in transgenic line B,and the least total adult emergence in the transgenic line C in F0 and F3 generations.Conclusions These findings suggest that the transgenic cotton lines expressing fern protein disrupts whitefly populations and the life cycle to a certain extent.However,results are not consistent over generations and years of study,indicating these transgenic lines were not superior over control lines and need to be improved in future breeding.
文摘The current study comprehensively evaluates four different protein extraction methods based on urea,sodium dodecyl sulfate(SDS),anionic surfactants(BT),and total RNA extractor(Trizol),aiming to optimize the sample preparation workflow for mass spectrometry-based proteomics.Using HeLa cells as an example,we found that the method employing the mass spectrometry-compatible surfactant BT reagent significantly reduces the total time consumed for protein extraction and minimizes protein losses during the sample preparation process.Further integrating the four protein extraction methods,we identified over 7000 proteins from HeLa cells without relying on pre-fractionation techniques,and 2990 of them were quantified using label-free quantification.It is worth noting that the BT and SDS methods demonstrate higher efficiency in extracting membrane proteins,while the Urea and Trizol methods are more effective in extracting proteins from nuclear and cytoplasmic fractions.In summary,this study provides a novel solution for deep proteome coverage,particularly in the context of cellular protein extraction,by integrating mass spectrometry-compatible surfactants with traditional extraction methods to effectively enhance protein identification numbers.
文摘Profiling the protein composition of bacteria is essential for understanding their biology,physiology and interaction with environment.Mass spectrometry has become a pivotal tool for protein analysis,facilitating the examination of expression levels,molecular masses and structural modifications.In this study,we compared the performance of three widely-used mass spectrometry methods,i.e.,matrix-assisted laser desorption/ionization(MALDI)protein fingerprinting,top-down proteomics and bottom-up proteomics,in the profiling of bacterial protein composition.It was revealed that bottom-up proteomics provided the highest protein coverage and exhibited the greatest protein profile overlap between bacterial species.In contrast,MALDI protein fingerprinting demonstrated superior detection reproducibility and effectiveness in distinguishing between bacterial species.Although top-down proteomics identified fewer proteins than bottom-up approach,it complemented MALDI fingerprinting in the discovery of bacterial protein markers,both favoring abundant,stable,and hydrophilic bacterial ribosomal proteins.This study represents the most systematic and comprehensive comparison of mass spectrometry-based protein profiling methodologies to date.It provides valuable guidelines for the selection of appropriate profiling strategies for specific analytical purposes.This will facilitate studies across various fields,including infection diagnosis,antimicrobial resistance detection and pharmaceutical target discovery.
基金This work was supported by the National Key Research and Development Program of China(2021YFA1301504)the Chinese Academy of Sciences Strategic Priority Research Program(XDB37040202)the National Natural Science Foundation of China(91953101).
文摘The assembly of a protein complex is very important for its biological function,which can be investigated by determining the order of assembly/disassembly of its protein subunits.Although static structures of many protein com-plexes are available in the protein data bank,their assembly/disassembly orders of subunits are largely unknown.In addition to experimental techniques for studying subcomplexes in the assembly/disassembly of a protein complex,computational methods can be used to predict the assembly/disassembly order.Since sampling is a nontrivial issue in simulating the assembly/disassembly process,coarse-grained simulations are more efficient than atomic simulations are.In this work,we developed computational protocols for predicting the assembly/disassembly orders of protein complexes via coarse-grained simulations.The protocols were illustrated via two protein complexes,and the predicted assembly/disassembly orders were consistent with the available experimental data.
文摘目的研究尘螨肠道微生物蛋白Hypothetical protein CE2118的表达和纯化,为研究其在尘螨疫苗免疫治疗中的作用奠定基础。方法采用生物信息学方法,根据GenBank中Hypothetical protein CE2118蛋白的基因序列,将其中稀有密码子改造为大肠杆菌常用密码子并进行二级结构优化,合成Hypothetical protein CE2118基因,构建原核表达载体pGEX6P-1-hypothetical protein CE2118并经酶切鉴定,在大肠埃希菌Rosetta(DE3)中用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,重组产物采用GST亲和层析纯化柱纯化。结果经密码子改造和二级结构优化后Hypothetical protein CE2118基因长度为588bp,其编码蛋白理论分子量为21kDa。重组表达载体经酶切鉴定与理论推测结果相符,该基因经IPTG诱导在大肠埃希菌Rosetta(DE3)中得到高效的可溶性表达,纯化后的重组蛋白分子量约为21kDa,其单一蛋白纯度达95%以上。结论本研究成功构建了Hypothetical protein CE2118基因的pGEX6P-1原核重组质粒,获得的可溶性重组蛋白为进一步研究肠道微生物蛋白Hypothetical protein CE2118在尘螨疫苗免疫治中的作用机理奠定基础。
基金Supported by Heilongjiang Province Science and Technology Key Project(GC04B115)
文摘In winter, spring and summer, the rhizome of wild Elytrzgia repens of Heilongjiang Province was selected to extract the soluble which whole protein and the apoplastic protein, and analyzed by SDS-PAGE. The result indicated that there were two specific polypeptides in two types protein from winter; their relative molecular weight were identified as 52 ku and 26 ku by analyzing software; the apoplastic protein from winter had the ability of modifing the growth of ice crystal which appeared hexagonal in shape observed with the phase-contrast photomicroscope. So the apoplastic protein from winter has the antifreeze characters and the 52 ku protein is more likely the antifreeze protein
文摘Background Increased levels of inflammatory markers have been documented in various settings of coronary artery disease. The vulnerability of coronary lesions in acute myocardial infarction(AMI) at the time of onset may be related to serum levels of C reactive protein(CRP) on admission, before CRP levels are affected by myocardial damage.Objective This study assessed the predictive value of CRP levels within six hours after the onset of acute anterior myocardial infarction with primary percutaneous coronary intervention(PCI).Methods The plasma CRP of 76 patients with first acute anterior myocardial infarction was measured within 6 hours after onset. They were divided into 2 groups: group 1( n =20) with elevated CRP( ≥0.3mg/dl ) on admission within 6 hours after onset and group 2( n =56) with normal CRP( <0.3mg/dl ) within 6 hours after onset. All patients were treated by primary PCI. The primary combined end points, including death due to cardiac causes, re MI related to the infarction artery(RIA) and repeat intervention of the RIA, and the restenosis rate were assessed in relation to CRP levels within 6 hours after onset. Left ventricular end diastolic volume index(EDVI),end systolic volume index(ESVI),and ejection fraction(EF) on admission and 6 month after the onset were assessed by left ventriculography. Changes in EDVI(ΔEDVI),ESVI(ΔESVI), and EF(ΔEF) were obtained by subtracting respective on admission values from corresponding 6 month follow up values. Results There were no significant differences in baseline characteristics between the two groups. The primary combined end points were significantly more frequent in group 1(20%) than those in group 2( 1.79% , P <0.01 ).In addition, restenosis rates were significantly higher in group 1 than in group 2(41.18% vs 16.07%, P<0.05). Group 1 showed greater increases in left ventricular volume and less improvement in EF compared with group 2(ΔEDVI 6.31 ±2.17 vs 3.29 ±9.46ml/m 2 , ΔESVI 5.92 ±2.31 vs 3.86 ±1.08ml/m 2 , ΔEF 1.92 ±0.47 vs 4.79 ±1.73% , P <0.05 , respectively).Conclusions CRP levels within 6 hours after the onset of AMI might predict adverse outcome after primary PCI and progressive ventricular remodeling within 6 month of AMI.
基金supported by the National Programs for High Technology Research and Development of China (2006AA10A204)the Gansu Key Technologies R&D Program(ZGS-052-A41-0006-03)the Programs for Director Fund of Lanzhou Veterinary Research Institute
文摘We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine.
基金Mega-Projects of S&T Research for the 10th Five- Year Plan from Ministry of S&T of the People’s Republic of China(2001BA501A02B) Shenzhen S&T Tri- item Foundation (S040145)
文摘This pilot study involved the application of a crossflow ultrafiltration （UF） membrane module employing hollow fiber polysulphone membranes in the reclamation of protein and oligosaccharides from soy protein wastewater. The optimal membrane molecular weight cut-off （MWCO） was selected as 10 ku upon retaining ratios of protein and oligosaccharides as well as the variation of permeate flux. The effects of pretreatment strategy and operating conditions, such as transmembrane pressure （TMP）, feed pH and velocity on flux were studied. According to the experimental result, the optimal operating conditions were determined as temperature 45 ℃, pH 4.5, TMP 0.2 MPa and feed velocity 10 L·s^-1. In addition, the permeate flux recovery by pure water backflushing and chemical cleaning was investigated. The experimental result showed that backflushing with pure water could only recover the lost permeate flux by 30%, and the decrease of backflushing interval was helpful in improving UF permeate productivity. Chemical cleaning test revealed that the combination of sodium hydroxide and EDTA was an ideal agent for cleaning the fouled membranes.
