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surface display of Rice Stripe Virus NSvc2 and Analysis of Its Membrane Fusion Activity

Virologica Sinica 2012年第2期27卷 100-108页

作者： Shu-ling Zhao Xue-juan Dai Jian-sheng Liang Chang-yong Liang College of Bioscience and Biotechnology Yangzhou UniversityYangzhou 225009China

Rice stripe virus (RSV) infects rice and is transmitted in a propagative manner by the small brown planthopper. How RSV enters an insect cell to initiate the infection cycle is poorly understood. Sequence analysis revealed that the RSV NSvc2 protein was similar to the membrane glycoproteins of several members in the family Bunyaviridae and might induce cell membrane fusion. To conveniently study the membrane fusion activity of NSvc2, we constructed cell surface display vectors for expressing Nsvc2 on the insect cell surface as the membrane glycoproteins of the enveloped viruses. Our results showed that NSvc2 was successfully expressed and displayed on the surface of insect Sf9 cells. When induced by low pH, the membrane fusion was not observed in the cells that expressed NSvc2. Additionally, the membrane fusion was also not detected when co-expressing Nsvc2 and the viral capsid protein on insect cell surface. Thus, RSV NSvc2 is probably different from the phlebovirus counterparts, which could suggest different functions. RSV might enter insect cells other than by fusion with plasma or endosome membrane.

关键词： Rice stripe virus NSvc2 surface display Membrane fusion

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surface display of OmpC of Salmonella serovar Pullorum on Bacillus subtilis spores

Surface display of OmpC of Salmonella serovar Pullorum on Ba...

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中国畜牧兽医学会动物微生态学分会第五届第十三次全国学术研讨会

作者： Xixi Dai Minggang Liu Kangcheng Pan Jinlong Yang Chongqing Academy of Animal Science College of Veterinary Medicine Sichuan Agricultural University

Salmonellosis is a major public health problem throughout the ***,there is a huge need for diversified control strategies for Salmonella *** this work,we have assessed the potential use of Bacillus subtilis(***) spores for the expression of a major protective antigen of Salmonella serovar Pullorum,*** expression of OmpC on the surface of spores was determined by immunofluorescence *** immunized with recombinant spores expressing the OmpC antigen presented significant levels of OmpC-specific serum IgG and mucosal SIgA antibodies than in mice immunized with non-recombinant spores(p<0.01).In addition,oral immunization with recombinant spores was able to induced a significant level of protection in mice against lethal challenge with Salmonella serovar *** results suggest that *** spores have promising potential in the development of mucosal vaccines against Salmonella infections.

关键词： Bacillus subtilis Spores Recombinant surface display Salmonella pullorum OmpC Mucosal vaccines

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surface display of a bacterial amylase enzyme on yeast

Surface display of a bacterial amylase enzyme on yeast

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International Conference on Food Engineering and Biotechnology（ICFEB 2011）

作者： Nadzarah A.Wahab Zainon Mohd Noor Mohamad Faiz Foong Abdullah Dept of Biomolecular Science Faculty of Applied Sciences Universiti Teknologi MARA Malaysia

An ***-yeast shuttle vector for the anchoring of heterologous protein to the yeast host’s cell wall was constructed using the backbone from pGAD424.A construct comprising the signal sequence from the yeast sucrose isomerase gene（SucSg）,a multiple cloning site sequence and a DNA fragment encoding 67 amino acids from the carboxyl-terminal of the yeast cell wall protein 2（CWP2） was constructed in *** construct was designed such that a gene sequence cloned into the MCS will be translated in-frame with the SucSg and *** construct was then inserted into the HindIII site on pGAD424,replacing the GAL4 fusion tag and the original MCS *** sequencing confirmed the correct insertion of both signal and anchor proteins in the vector.A bacterial amylase gene was cloned into the vector and tested for functionality.

关键词： surface display bacterial amylase yeast pGAD424.

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display of aggregation-prone ligand binding domain of human PPAR gamma on surface of bacteriophage lambda

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Acta Pharmacologica Sinica 2006年第1期27卷 91-99页

作者： Bo KONG~(2,4) Wei-jun MA~(2,3,5)2 Institute of Health Sciences.Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences and Medical School of Shanghai Jiao Tong University.Shanghai 200025.China 3 Shaughai Research Center of Biotechnology,Shanghai Institutes for Biological Sciences,Chiese Academy og Sciences,Shanghai 200233.China 4 Graduate School of the Chinese Academy of Sciences Institute of Health Sciences Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Medical School of Shanghai Jiao Tong University Shanghai China Graduate School of the Chinese Academy of Sciences China Shanghai Research Center of Biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Aim:To display the aggregation-prone ligand binding domain (LBD) of the hu-man peroxisome proliferator-activated receptor gamma (PPARγ) on the surface ofbacteriophages to establish an easy screening assay for the identification of PPARγ***:Plasmids were constructed for the expression of the PPARγLBD as a fusion to the N-terminus of the g3p protein of filamentous phage or theC-terminus of the capsid protein D (pD) of phage *** fusion proteinswere expressed in E coli and solubility characteristics were *** against the LBD as well as the pD protein were prepared for Westernblot analysis and phage capture ***:The pD-LBD fusion protein waspartially soluble,whereas the LBD-g3p fusion protein was detected only in theinsoluble *** pD-LBD fusion protein was efficiently incorporated inphage ***,the LBD was shown to be displayed on the surfaceof bacteriophage *** average,the pD-LBD fusion protein accounted for28% of the total pD protein in the lambda head ***:The hydro-phobic PPARγ LBD was expressed as a soluble form of fusion protein in E coli anddisplayed on the surface of bacteriophage lambda when it was fused to the lambdapD *** lambda pD fusion system could be used for improving the solu-bility of proteins that tend to form inclusion bodies when expressed in E *** phage particles displaying the LBD of PPARy may be of great value for theidentification of novel PPARγ ligands.

关键词： protein expression lambda phage surface display nuclear receptor peroxisome proliferator-activated receptors ligand binding domain

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Whole‑cell catalytic synthesis of trehalose by Corynebacterium glutamicum displaying trehalose synthase on its cell surface

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Systems Microbiology and Biomanufacturing 2023年第3期3卷 489-497页

作者： Ming Fang Qiang Wang Zhi Wang Ruiqi Lu Rufan Xing Xian Zhang Zhiming Rao Key Laboratory of Industrial Biotechnology of Ministry of Education School of BiotechnologyJiangnan UniversityWuxi 214122Jiangsu ProvincePeople’s Republic of China

Trehalose is a disaccharide with many applications in cosmetics,refrigeration,and *** synthase is a significant enzyme in trehalose *** coli is usually used to express this enzyme *** *** is a pathogenic strain,we chose Corynebacterium glutamicum ATCC13032 as an engineering strain in this study for food safety *** of its poor permeability,we constructed two recombinant *** strains using two anchor proteins,PorH,and short-length NCgl1337,to anchor trehalose synthase from Streptomyces coelicolor on the cell surface and synthesize trehalose directly from *** on enzymatic properties indicated that NCgl1337S–ScTreSK246A had better enzyme activity and thermal stability than the free *** optimizing the whole-cell transformation,the optimal transformation condition was 35°C,pH 7.0,and OD600 of *** this condition,the conversion rate of 300 g/L maltose reached 69.5%in a 5 L *** relative conversion rate was still above 75%after repeated five times.

关键词： Trehalose synthase Corynebacterium glutamicum surface display Whole-cell catalyses

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