Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Chinese Journal of Cellular and Molecular Immunology》 2015-01
Add to Favorite Get Latest Update

Preparation and identification of the polyclonal antibody against heat shock protein 70(DnaK) from Shigella flexneri 5a M90T

WANG Yu;ZHOU Wei;LIAO Xiang;YUE Jun Jie;SONG Ting;DAI Hongmei;CHEN Xinxin;LIANG Long;Huhe Bateer;College of Veterinary Medicine,Inner Mongolia Agricultural University;Beijing Institute of Biotechnology,Academy of Military Medical Sciences;  
Objective To generate polyclonal antibodies against Shigella flexneri 5a M90 T heat shock protein 70(Dna K)and identify the specificity and titer of antibodies.Methods The sequence of Shigella flexneri 5a M90 T dna K gene was amplified by PCR and inserted into expression vector p ET-24 a to obtain recombinant plasmid p ET24a-dna K.Then the recombinant plasmid p ET24a-dna K was transformed into E.coli BL21(DE3) and expressed under IPTG induction.The recombinant Dna K proteins with His-tag(Dna K-His) were then purified by affinity purification.The polyclonal antibodies were prepared by immunizing the mice with the purified recombinant proteins and then harvesting mouse sera.The specificity and titer of the polyclonal antibodies were identified by Western blotting,ELISA and immunofluorescent staining.Results The recombinant plasmid p ET24a-dna K was expressed efficiently in E.coli BL21(DE3).The polyclonal antibodies harvested from the mice immunized with Dna K-His were demonstrated being specific to Shigella flexneri 5a M90 T Dna K proteins,and could be efficient in Western blotting,ELISA and immunofluorescent staining.Conclusion The polyclonal antibodies against Shigella flexneri 5a M90 T Dna K protein have been successfully prepared.
【Fund】: 国家自然科学基金(30970122)
【CateGory Index】: R392
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
【Citations】
Chinese Journal Full-text Database 2 Hits
1 Gong Yixiong1 Liao Xiang2 Zhu Li2 Wu Lan1 Zhou Xiaowei2 Liang Long2 Huang Peitang2(1School of Life Sciences and Food Engineering,Nanchang University,Nanchang 330047; Institute of Biotechnology,Academy of Military Medical Science,Beijing 100071);Identification of Virulence-related Membrane Protein Complex from Shigella flexneri 5a M90T by BN/SDS-PAGE[J];Biotechnology Bulletin;2011-09
2 LIU Ying, ZHU Mei-cai, WANG Yin-jing, ZHAN Zhi, LIU Zheng-gang Center for Clinical Molecular Biology Research, Airforce General Hospital of Chinese PLA, Beijing 100036, China;Cloning of a DnaJ homolog chaperon PBP and its subcellular localization[J];Journal of Cellular and Molecular Immunology;2003-06
【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 WANG Shu-mao;LAN Dao-liang;CHEN Ya-bing;LIN Bao-shan;LI Jiang;College of Life Science and Technology,Southwest University for Nationalities;College of Tibetan Plateau Research,Southwest University for Nationalities;;Detection of Tissue Distribution of HSPA1A and HSPA2 Genes in Yaks[J];Progress in Veterinary Medicine;2013-12
2 Lian Dongmei;Wang Huifang;Chen Miancai;Environment and Plant Protection College,Hainan University;Institute of Environment and Plant Protection,Hainan Academy of Agricultural Sciences;;Research Progress on Hsp70 of Plant Parasitic Nematodes[J];Genomics and Applied Biology;2013-06
3 XIA Run-lin;PAN Hou-jun;ZHAO Fei;FANG Xiang;SHI Cun-bin;LI Hua;WU Shu-qin;College of Fisheries and Life Science,Dalian Ocean University;Key Laboratory of Fishery Drug Development,Ministry of Agriculture,Key Laboratory of Aquatic Animal Immune Technology,Guangdong Province,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences;;Cloning and expression analysis of HSP70 ORF in Ichthyophthirius multifiliis[J];Journal of Dalian Ocean University;2014-04
4 Pang Xin;Chen Yunzhi;College of Packaging and Printing Engineering,Tianjin University of Science & Technology;;Study of Antibacterial Properties of Nanometer ZnO[J];Packaging Journal;2014-04
5 XIA Huan;ZHANG Qi-zhong;Institute of Hydrobiology,Jinan University/Key Laboratory of Eutrophication and Red Tide Prevention of Guangdong Higher Education Institutes;;Response of HSP70s in Crassostrea hongkongensis to a Pollutant Nonylphenol[J];Journal of Ecology and Rural Environment;2014-02
6 ZHANG Jing;HE Ning;ZHANG Da-long;ZHOU Peng-hui;GUAN Tong;JIANG Shu-qing;ZHANG Jing-shu;Department of Toxicology,Tianjin Centers for Disease Control and Prevention;;Expression of heat shock protein 70 and anti-oxidant effect induced by lead nitrate in L02 cell line[J];Journal of Environment and Health;2014-02
7 XIE Yanhai;YU Zexi;WEN Chungen;HU Baoqing;Department of Medical Laboratory Animal Science,Nanchang University;Jiangxi Fisheries Research Institute;Department of Bio-science,Nanchang University;;Prokaryotic expression and preparation of polyclonal antibodie of heat shock protein 70 in Cristaria plicata[J];Journal of Nanchang University(Natural Science);2014-03
8 WANG Ling;YANG Shuai;ZHU Ming-He;XU Zhong-Xin;ZHAO Kui-Jun;HAN Lan-Lan;College of Agriculture,Northeast Agricultural University;Virus-Free Seedling Research Institute,Heilongjiang Academy of Agricultural Sciences;;Analysis of transcription differences of the heat shock cognate 70 gene in different tissues in response to heat stress in Xestia c-nigrum Linnaeus(Lepidoptera: Noctuidae)[J];Chinese Journal of Applied Entomology;2014-04
9 WANG Qiang;LEI Yan;LI Min;WANG Yang;Experimental Research Center,China Academy of Chinese Medical Sciences;Guang'anmen Hospital,China Academy of Chinese Medical Sciences;;Relationship and Research Progress between Heat Shock Protein 27 and Cardiovascular Disease[J];Liaoning Journal of Traditional Chinese Medicine;2014-09
10 MAO Pei;DU Meng-Fang;YAO Shuang-Yan;CAI Yong-Ping;GUO Xian-Ru;YUAN Guo-Hui;LUO Mei-Hao;College of Plant Protection,Henan Agricultural University;;Expression profiling of salivary gland proteins and the corresponding genes in Helicoverpa armigera( Lepidoptera: Noctuidae) larvae feeding on artificial diets containing fruits of different pepper varieties[J];Acta Entomologica Sinica;2014-10
【Secondary Citations】
Chinese Journal Full-text Database 1 Hits
1 LIU Cheng gang, ZHU Mei cai, Zhan Zhi, et al (Center of Clinical Molecular Biology, General Hospital of Air Force, Beijing 100036, China);Improvement of luciferase refolding in vitro by peripherin-binding protein of photoreceptor cell[J];Basic Medical Sciences and Clinics;2002-06
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved