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| Prokaryotic Expression of Porcine FcγRⅢ and Production of Its Polyclonal Antibody |
| XU Hong-yun,XIA Ping-an,ZHANG Feng-hua,ZHAO Lin,FAN Xu,LIU Ming-li,LIU Yu-song,ZHANG Zhi-yuan,CUI Bao-an |
| The College of Animal Husbandry and Veterinary,Henan Agricultural University, Zhengzhou 450002, China |
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Abstract Objective:To construct a prokaryotic plasmid expressing the recombinant proteins of porcine FcγRIII and prepare the polyclonal antibodies of mouse anti porcine FcγRⅢ. Methods The fragment of gene encoding porcine FcγRIII was amplified from pTG19-T-FcγRIII plasmid by PCR. The fragment was then inserted into the prokaryotic expression vector pET-32a to construct the recombinant plasmid pET-FcγRIII, and expressed in E. coli BL21 (DE3). After induction with IPTG, the fusion protein was obtained , and then purified with Urea by washing . A polyclonal antibody , analyzed by Western blotting and ELISA staining , was developed by immunizing mouse with the purified recombinant protein. Results: The recombinant plasmid was constructed successfully ; the fusion protein was successfully expressed and purified. Western blotting and ELISA staining demonstrated that the polyclonal antibody was obtained by immunizing mouse with the purified recombinant protein. The results of ELISA and Western blotting indicated that the prepared polyclonal antibody had high titer and specificity.Conclusion: The preparation of the polyclonal antibody against porcine FcγRIII lays a foundation for the further study on the function of porcine FcγRIII protein.
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Received: 14 December 2009
Published: 12 June 2010
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Cite this article:
XU Gong-Yun, JIA Beng-An, ZHANG Feng-Hua, DIAO Lin, FAN Xu, LIU Meng-Chi, LIU Yu-Song, ZHANG Zhi-Yuan, CUI Bao-An. Prokaryotic Expression of Porcine FcγRⅢ and Production of Its Polyclonal Antibody. China Biotechnology, 2010, 30(06): 117-121.
URL:
https://manu60.magtech.com.cn/biotech/Q819 OR https://manu60.magtech.com.cn/biotech/Y2010/V30/I06/117
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