研究简报 Short Communications

鸡β-防御素-9的融合表达及其多克隆抗体的制备与鉴定

  • 许兰娇 ,
  • 洪智敏 ,
  • 司微 ,
  • 刘宝生 ,
  • 张锦华 ,
  • 刘思国 ,
  • 黎观红
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  • 1. 江西农业大学动物科技学院,南昌 330045;
    2. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室,哈尔滨 150001

收稿日期: 2012-09-01

  网络出版日期: 2013-02-28

基金资助

国家自然科学基金项目(30800791);江西省青年科学家培养对象计划项目(赣科发计字[2010]209号);江西省教育厅科技项目(GJJ10410)

Fusion Expression of Chicken β-Defensin-9 and Preparation, Characterization of Its Polyclonal Antibody

  • XU Lanjiao ,
  • HONG Zhimin ,
  • SI Wei ,
  • LIU Baosheng ,
  • ZHANG Jinhua ,
  • LIU Siguo ,
  • LI Guanhong
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  • 1. College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China;
    2. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China

Received date: 2012-09-01

  Online published: 2013-02-28

摘要

为制备鸡β-防御素-9(AvBD9)多克隆抗体,本试验将AvBD9基因插入pGEX-6P-1载体,构建重组基因pGEX6P-AvBD9大肠杆菌表达质粒。将通过切胶纯化获得的表达产物包涵体谷胱甘肽转移酶-AvBD9(GST-AvBD9)融合蛋白免疫新西兰大白兔,制备抗AvBD9血清,并检测抗体效价。结果表明,经间接酶联免疫分析(ELISA)及蛋白质免疫印迹(Western blot)方法证实,获得的融合蛋白免疫新西兰大白兔得到了特异性的多克隆抗体,抗体效价为1:38 400。

本文引用格式

许兰娇 , 洪智敏 , 司微 , 刘宝生 , 张锦华 , 刘思国 , 黎观红 . 鸡β-防御素-9的融合表达及其多克隆抗体的制备与鉴定[J]. 动物营养学报, 2013 , 25(3) : 603 -608 . DOI: 10.3969/j.issn.1006-267x.2013.03.019

Abstract

To prepare polyclonal antibody against chicken β-defensin-9 (AvBD9), AvBD9 gene was cloned into the prokaryotic expression vector pGEX-6P-1. The recombinant expression plasmid pGEX-6P-AvBD9 was constructed and then transformed into Escherichia coli (E. coli) to express AvBD9. The glutathione transferase-AvBD9 (GST-AvBD9) fusion protein was expressed in E. coli as an inclusion body. The fusion protein purified from the PAGE gel was used to immunize New Zealand white rabbit. The results showed that as detected by indirect ELISA and Western blot, polyclonal antibody against AvBD9 with the titer of 1:38 400 was obtained from the immunized rabbits.

参考文献

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