检索结果-南通市图书馆

fusion expression of immunological epitopes from VP1-3 of foot-and-mouth disease virus with porcine interferon-gamma

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Fusion expression of immunological epitopes from VP1-3 of fo...

2006年度学术研讨会

作者： YAO Qing-xia, QIAN Ping, CAO Yi, XU Zhuo-fei, HE Yan-nan, SI You-hui, CHEN Huan-chun (Laboratory of Animal Virology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei Province, 430070, China)

The artificial synthetic immunological epitopes from VP1-3 of foot-and-mouth disease virus (FMDV) were inserted into prokaryotic expression vector (pGEX-KG) in cascade connection with porcine interferon-γ. The recombinant plasmids were identified by enzymes and sequencing. The identified recombinant plasmid was transformed into BL21 and induced by IPTG to express the fusion protein in high quantities. The expressed product was analyzed by SDS-PAGE, and the results demonstrated that the molecular weight of it was about 66 kDa, which was consisted with the anticipation. And the interest protein was about 37.4% of the total protein by scanning, which mainly existed in inclusion bodies. After these inclusion bodies were disnatured by SKL, they were renatured by PEG4000, oxidation tathion and reduction tathion. The crude extracted product was analyzed by Western-blotting, which indicated that the expressed protein had the biologic activity. This study established foundation to the next work of genetically engineered vaccines.

关键词： foot-and-mouth disease virus antigen epitope porcine interferon-γ fusion expression

Eukaryotic Secrete fusion expression of Rhizopus oryzae glucoamylase and Paenibacillus sp.xylanase

在线全文

cnki会议

学校读者我要写书评

暂无评论

Eukaryotic Secrete fusion expression of Rhizopus oryzae gluc...

第五届全国微生物资源学术暨国家微生物资源平台运行服务研...

作者： Wang Zhi Dun Bao-Qing Lu Ming The National Key Facility for Crop Gene Resources and Genetic Improvement of Institute of Crop Sciences Chinese Academy of Agricultural Sciences

The glucoamylase gene(amy A) was amplified from Rhizopus oryzae 3.866 total DNA extracts. The result showed that the sequence of amyA was consisted of 2049 bp,and encoded 604 amino acids.The xylanase gene(xynA) was amplified from Paenibacillus sp.H10-3 total DNA extracts.The result showed that the mature peptide sequence of xynA was consisted of 636bp,and encoded 211 amino acids.Then,the mature peptide sequence of amyA gene,sequence of linker and sequence of xyn A gene were spliced by overlap extension PCR(SOE-PCR),which amyA-l-xynA was obtained.The fusion gene was then inserted into a Pichia pastoris expression vector pPIC9 to produce the recombinant expression plasmid p?lC9-amyA-l-xynA.By using electric shocks,we successfully transformed the recombinant pP\C9-amyA-l-xynA into Pichia pastoris GS115,which AX 11 was obtained..The maximum yield of the recombinant glucoamylase and xylanase in AX11 culture medium were 5.8U/mL and 32.3U/mL, respectively.

关键词： glucoamylase xylanase fusion expression Pichia pastoris

Engineering Saccharomyces cerevisiae for enhanced(–)-α-bisabolol production

在线全文

cnki会议

学校读者我要写书评

暂无评论

Synthetic and Systems Biotechnology 2023年第2期8卷 187-195页

作者： Yinkun Jiang Lu Xia Song Gao Ning Li Shiqin Yu Jingwen Zhou Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology Jiangnan University1800 Lihu RoadWuxiJiangsu214122China Science Center for Future Foods Jiangnan University1800 Lihu RoadWuxiJiangsu214122China Key Laboratory of Industrial Biotechnology Ministry of Education and School of BiotechnologyJiangnan University1800 Lihu RoadWuxiJiangsu214122China Jiangsu Province Engineering Research Center of Food Synthetic Biotechnology Jiangnan UniversityWuxi214122China

(–)-α-Bisabolol is naturally occurring in many plants and has great potential in health products and pharma-ceuticals.However,the current extraction method from natural plants is unsustainable and cannot fulfil the increasing requirement.This study aimed to develop a sustainable strategy to enhance the biosynthesis of(–)-α-bisabolol by metabolic engineering.By introducing the heterologous gene MrBBS and weakening the competitive pathway gene ERG9,a de novo(–)-α-bisabolol biosynthesis strain was constructed that could produce 221.96 mg/L(–)-α-bisabolol.Two key genes for(–)-α-bisabolol biosynthesis,ERG20 and MrBBS,were fused by a flexible linker(GGGS)3 under the GAL7 promoter control,and the titer was increased by 2.9-fold.Optimization of the mevalonic acid pathway and multi-copy integration further increased(–)-α-bisabolol production.To promote product efflux,overexpression of PDR15 led to an increase in extracellular production.Combined with the optimal strategy,(–)-α-bisabolol production in a 5 L bioreactor reached 7.02 g/L,which is the highest titer reported in yeast to date.This work provides a reference for the efficient production of(–)-α-bisabolol in yeast.

关键词： Metabolic engineering (–)-α-bisabolol fusion expression Transporter Saccharomyces cerevisiae

Overexpression of Soluble Human Thymosin Alpha 1 in Escherichia coli

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Acta Biochimica et Biophysica Sinica 2005年第2期37卷 147-151页

作者： Pei-Fu CHEN Hong-Ying ZHANG Geng-Feng FU Gen-Xing XU Ya-Yi HOU Medical School,Nanjing University.Nanjing 210093,China School of Life Sciences.Nanjing University.Nanjing 210093,China Medical School Nanjing University Nanjing 210093 China School of Life Sciences Nanjing University Nanjing 210093 China

Synthesized gene of human thymosin alpha 1（Tα1）was inserted into pET-28a,pET-9c,pThioHis B.pGEX-2T or pBV222 and then inductively expressed in strains of Escherichia coli.Among thefive expression systems,the BL21/pET-28a system provides the highest expression level of fusion protein ina soluble form,which is up to 70% of total expressed bacterial proteins as visualized by sodium dodecylsulfate-polyacrylamide gel electrophoresis（SDS-PAGE）.The resulting fusion protein purified through nickelaffinity chromatography accounts for 2.53% of the wet bacterial pellet weight and reaches 94.5% purity bySDS-PAGE.These results indicate the potential of this expression system for high-throughput production ofrecombinant Tα1.

关键词： human thymosin alpha l Escherichia coli fusion expression Ni2+ affinity chromatography

Prokaryotic expression of Antimicrobial Peptide CATH PR1–2 from the Skin of Paa robertingeri in Escherichia coli

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Asian Herpetological Research 2017年第4期18卷 275-283页

作者： Huaiqing DENG Chen CHEN Ning XIAO Jiang ZHOU College of Life Sciences Guizhou Normal University Guiyang 550001 Guizhou China

The aim of this study was to investigate the prokaryotic expression of antimicrobial peptide cathelicidin （CATH） PR1 and PR2 from the skin of Paa robertingeri in Escherichia coli. Two active peptides, CATH PR1 and CATH PR2, belong to the CATH family in the skin of P. robertingeri. CATH PR1 has a relatively high antimicrobial activity, especially for the drug-resistant strains found in clinical practice; however, no antimicrobial activity has been found in CATH PR2. The molecular weights of both CATH PR1 and CATH PR2 are relatively low （3195.88 and 2838.34 Da, respectively）. Thus, the genetic processes, as well as the expression and purification of these proteins, are difficult to perform. Therefore, in this study, CATH PR1 and CATH PR2 genes were tandem ligated and then connected to the plasmid pET-32a. This reconstructed plasmid was then transfected into the expression vector E. coli BL21 to construct the recombinant expression system. The fusion expression of peptide PR was stable in E. coli after induction with 1.0 mol/L isopropyl β-D-1-thiogalactopyranoside at 37℃ for 4 h. The antimicrobial activity assay using Staphylococcus aureus （Song） and Candida albicans 08030102 showed that the antimicrobial activity of PR was similar to the antimicrobial activity of CATH PR1. This study showed that artificial modification of the amino acid sequences of PR1 and PR2 could result in better protein expression in prokaryotes, and the fusion protein expressed had relatively high antimicrobial and other biological activities. In conclusion, the findings suggest future prospects of the commercialization of this method.

关键词： E. coli BL21 fusion expression Paa robertingeri recombinant protein PR

Advances in Antimicrobial Peptides and expression Strategy

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Journal of Northeast Agricultural University(English Edition) 2011年第1期18卷 85-90页

作者： SUN Yan WANG Yunfeng CHEN Hongyan SHI Xingming WANG Mei HUANG Tingting SUN Peng YANG Guihua College of Life Sciences Northeast Agricultural University Harbin 150030 China Division of Avian Infectious Diseases National Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences Harbin 150001 China

Antimicrobial peptides are widely distributed in nature,existing in organisms of plants,insects,and vertebrates.It has been approved that antimicrobial peptides have broad spectrum antimicrobial activities,and play a key modulatory role in the innate immune response and tumor inhibiting activity.Due to the special action mechanism,the antimicrobial peptides become a hot field of genetic engineering.In the present paper,the general properties,mechanism of action,application value,existing problems,the latest progress and the expression strategy were discussed.

关键词： antimicrobial peptide molecular biology fusion expression

Production of Rabbit Polyclonal Antibody Using hAPOA1 Protein Expressed in Prokaryotic Cells

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Agricultural Biotechnology 2017年第2期6卷 41-44页

作者： Zhiwei XU Jie ZHAO Weizhong LIU Zhaoting LIU Qi WANG Zhenjiang Institute of Agricultural Sciences in Hilly Areas of Jiangsu Province Jurong 212402 China College of Life Science Nanjing Normal University Nanjing 210046 China

The open reading frame （ORF） of hAPOA1 was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX- 4T-I-hAPOA1, which was then transformed into Escherichia coil strain BL21. The expression of target fusion protein was induced with isopropyl β-D-l-thiogalacto- pyranoside （IPTG）. The purified fusion protein in inclusion bodies was used to immunize New Zealand white rabbits to prepare hAPOA1 antiserum and the antibody titer was detected with indirect enzyme-linked immunosorbent assay （ID-EL1SA）. ID-ELISA and Western Blot proved that rabbit polyclonal antibody with a high titer of 1 ： 40 000 was produced, which may bring considerable economic benefits.

关键词： HAPOA1 Prokaryotie expression New Zealand white rabbits fusion expression Polyelon~ antibody

Study of high-level expression of Musca domestica cecropin with partner molecular in Escherichia coli

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Study of high-level expression of Musca domestica cecropin w...

格莱姆抗菌肽——抗菌肽开发与应用技术研讨会

作者： Wang wei,Zheng Xueli*(Department of Pathogenic Biology,School of Public Health and Tropical Medicine,Southern Medical University,Guangzhou 510515,China)

Objective To utilize partner molecular for effective expressing Musca domestica cecropin,research that partner molecular influences expression of M.domestica cecropin in E.coli. Methods Gene of mature cecropin and ubiquitin of M.domestica were amplified by RT-PCR.Using bioinformatics methods,we analyzed secondary structure of the fusion protein of thioredoxin-MC and thioredoxin-ubiquitin-MC’s mRNA.Two expression plasmids of pET32a-MC and pET32a-Ubiquitin -MC were cloned.The expression plasmids were transformed into E.coli and were induced with IPTG respectively.Analysis of protein expression of pET32a-MC and pET32a-Ubiquitin-MC by SDS-PAGE was finished Results The protein quantity of expression of pET32a-Ubiquitin&MC were significantly higher than that of pET32a-MC.Conclusion Profile of mature peptide of cecropin was transformed by thioredoxin.The fusion protein of thioredoxin-MC isn’t toxic for host bacteria.The results showed the partner molecule ubiquitin can significantly result in high-level expression of peptide antibiotic of M. demostica cecropin.

关键词： Musca domestica Cecropin Partner molecular fusion expression Ubiquitin

Study on expression,Biological Stability and Anti-inflammatory Properties of a Novel Antimicrobial peptide Lacticin Q

在线全文

cnki会议

学校读者我要写书评

暂无评论

Study on Expression,Biological Stability and Anti-inflammato...

中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术...

作者： Lulu Zhang Dayong Si Xubiao Wei Rijun Zhang Laboratory of Feed Biotechnology State Key Laboratory of Animal NutritionCollege of Animal Science and TechnologyChina Agricultural University

The antimicrobial peptide Lacticin Q(LQ),which contains 53 amino acid residues,is a novel antimicrobial(AMPs) produced by Lactococcus lactis QU 5.LQ shows excellent thermostablity and acid-alkali tolerance and exhibits inhibitory activity against a wide range of pathogens.However,the low quantity of LQ produced by natural strains limited the further research and therapeutic application in feed industry.Recently,several biological systems have been developed in bacterial system for the high-production of AMPs.In our study,we efficiently expressed recomobinant LQ in Pichia pastoris(P.pastoris),and 32 mg pure LQ was obtained from fermentation liquid.The effects of temperature and pH on recombinant LQ was tested,and results showed that pH(from 3.0 to 8.0) and temperature(70,80,90 ℃) had a little effect on the biological activity of recombinant LQ.Another experiment was conducted to evaluate the effects of LQ on inflammatory response of mouse undergoing Staphylococcus aureus challenge.The results indicated that a low concentration of LQ(15 mg/mL) significantly reduced S.aureus-indaced inflammatory injury.Specifically,15 mg/mL LQ significantly decreased the decreased the expression of IL-1β,IL-6 and TNF-α(P < 0.05),which showed a direct activation of the inflammatory response that caused tissue destruction and a disease.In sum,this research not only supplied an effective approach for overproducing antibacterial peptide LQ,but paved the way for its further exploration on pharmaceutical potential and medical importance.

关键词： antimicrobial peptide Lacticin Q fusion expression Pichia pastoris antibacterial activity stablity