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Prokaryotic Expression of Rubber Elongation Factor Gene and Preparation of Its Polyclonal Antibody
Authors:CHEN Zhang-quan  WU Kun-xin  LIANG Xiao-dong  HUANG Zhen  LU Tian-tian
Affiliation:CHEN Zhang-quan1,WU Kun-xin2,LIANG Xiao-dong1,HUANG Zhen1,LU Tian-tian1 1.Department of Clinical Immunology,Guangdong Medical College,Zhanjiang 524023,2.Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou 571101
Abstract:[Objective] The aim of this study was to prepare the recombination protein of rubber elongation factor and its polyclonal antibodies.[Method] The encoding gene of rubber elongation factor(REF)was amplified by RT-PCR,and cloned into the prokaryotic expression vector pDEST17 to transform into Escherichia coil BI2I-AI.The recombinant protein induced by L-Arabinose was purified by the affinity chromatography.As the immunogen,the recombination protein was used to immunize mice for preparing polyclonal antibodies,while ELISA and Western blot hybridization were used to detect the titers and specificity.[Result] The purified recombination protein of REF with high expression was used to immunize house mice for preparing polyclonal antibodies with high titer and specificity.The western blot hybridization showed that the antibody could recognize the natural REF from latex.[Conclusion] The recombination protein of REF was successfully obtained and the mouse anti REF antibody with high titer and specificity was prepared,which lays a basis for further studies on biological functions of rubber elongation factor and other membrane proteins in rubber particles.
Keywords:Rubber elongation factor  Gene  Expression  Antibody  Preparation
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