鸡贫血病毒<em>VP1</em>基因的克隆及其在大肠杆菌中的表达

doi:10.3321/j.issn:1000-7091.2006.05.013

华北农学报 ›› 2006, Vol. 21 ›› Issue (5): 50-53. doi: 10.3321/j.issn:1000-7091.2006.05.013

所属专题： 畜牧；生物技术；

鸡贫血病毒VP1基因的克隆及其在大肠杆菌中的表达

黄冬艳¹, 杨兵², 齐欣³, 陈小玲², 徐福洲²

1. 江西农业大学动物科学院, 江西南昌 330045;
2. 北京市农林科学院畜牧兽医研究所, 北京 100089;
3. 沈阳农业大学畜牧兽医学院, 辽宁沈阳 110161

收稿日期:2006-03-12 出版日期:2006-10-28
通讯作者: 杨兵
作者简介:黄冬艳（1975- ），男，江西湖口人，讲师，主要从事兽医微生物教学与科研工作。
基金资助:
北京市青年创新基金项目（2005-2007）

Cloning of VP1 Gene of Chicken Infectious Anemia Virus and Its Expression in E. coli

HUANG Dong-yan¹, YANG Bing², QI Xin³, CHEN Xiao-ling², XU Fu-zhou²

1. College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China;
2. Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100089, China;
3. College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China

Received:2006-03-12 Published:2006-10-28

摘要/Abstract

摘要： 根据鸡贫血病毒Cux₁株基因组序列设计了一对引物，通过PCR扩增出VP1基因。将扩增出的片段克隆到pGM_T载体上，通过测序分析证实该片段与鸡贫血病毒Cux₁株的VP1基因序列一致。然后将克隆化VP1基因亚克隆到PET₃2a（+）载体上并进行原核表达。SDS_PAGE和Western Blot分析表明，一个分子质量为70.4 kDa的重组蛋白获得了成功表达，并且鸡贫血病毒阳性血清能和重组蛋白发生反应。

关键词: 鸡贫血病毒, VP1基因, 克隆, 表达

Abstract: We designed a pair of oligonucleotide primers based on the genomic sequence of CAV Cux_1 strain,and amplified VP1 gene by PCR.The amplification fragment was cloned into pGM_T easy vector.The result of nucleotide sequence analysis demonstrated that the cloned genomic DNA was identical with that deduced from Cux_1 strain of CAV.Then the cloned genomic DNA was subcloned into prokaryotic expression vector PET_32a(+),and the recombinant protein was expressed in E.coli cell.SDS_PAGE and Western Blot analysis showed that a protein with a molecular weight of 70.4 kDa was successfully expressed and CAV_infected chicken serum reacted with the recombinant protein.

Key words: Chicken infectious anemia virus, VP1, Cloning, Expression, E.coli

中图分类号:

S188

黄冬艳, 杨兵, 齐欣, 陈小玲, 徐福洲. 鸡贫血病毒VP1基因的克隆及其在大肠杆菌中的表达[J]. 华北农学报, 2006, 21(5): 50-53. doi: 10.3321/j.issn:1000-7091.2006.05.013.

HUANG Dong-yan, YANG Bing, QI Xin, CHEN Xiao-ling, XU Fu-zhou. Cloning of VP1 Gene of Chicken Infectious Anemia Virus and Its Expression in E. coli[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2006, 21(5): 50-53. doi: 10.3321/j.issn:1000-7091.2006.05.013.

http://www.hbnxb.net/CN/Y2006/V21/I5/50

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鸡贫血病毒VP1基因的克隆及其在大肠杆菌中的表达

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