鸡传染性法氏囊病病毒VP2基因密码子的优化及其在大肠杆菌中的表达  

Codon optimization of VP2 gene of infectious bursal disease virus and its expression in Escherichia coli

作　　者：潘群兴[1] 朱向东[1,2] 欧阳伟[1] 王晓丽[1] 夏兴霞[1] 毕振威[1] 诸玉梅[1] 董晨红[1] 王永山[1] 

机构地区：[1]江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室国家兽用生物制品工程技术研究中心,江苏南京210014 [2]南京农业大学动物医学院,江苏南京210095

出　　处：《中国兽医科学》2013年第3期271-276,共6页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31272537);江苏省自然科学基金项目(BK2010471;BK2012787);江苏省农业科技自主创新基金项目[cx(10)426]

摘　　要：为了提高鸡传染性法氏囊病病毒(IBDV)VP2基因在大肠杆菌中的表达量,分析近期引起安徽免疫失败IBD病例中分离的强毒分离株AH1株的VP2基因,发现有49个稀有密码子,其中编码Arg(R)的有16个,编码Gly(G)的有6个。编码精氨酸的AGG(AGA)2个稀有密码子紧密相连(R区),另外还有2个相对比较集中的GGG密集区(G区)。根据大肠杆菌密码子使用频率对IBD强毒分离株(AH1株)VP2基因进行改造并合成优化基因,命名为mVP2。将mVP2基因分别克隆到pET-32a及pBV220原核表达质粒,构建了含密码子优化型IBDV VP2基因的原核表达质粒pET32-VP2和pBV220-VP2。SDS-PAGE结果显示,优化基因mVP2在大肠杆菌中的表达较野生型基因有十分显著的提高。Western-blot检测到目的蛋白能够与鸡抗IBDV超免疫血清反应。本研究为开展VP2基因结构与功能的深入研究及IBD免疫预防和快速诊断奠定了理论基础。To improve the protein expression of VP2 of infectious bursal disease virus（IBDV） in Esc- herichia coli,VP2 gene structure of IBDV isolated from chicken with immunoprophylaxis defeat in Anhui was analyzed. There were 49 rare codons in the VP2 gene by contrast with codon usage in Escherichia coli, including 16 AGG/AGA（Arg） and 6 GGG（Gly）. Two AGG were joined together（called box R）and some GGG codons were relative concentrate（called box G）. The codon usage of wild type IBDV VP2 was opti- mized according to the codon bias of E. coli, and the full length optimized VP2 gene was synthesized, named mVP2,which was later cloned into the prokaryotic expression vectors pET-32a and pBV220 to construct recombinant expression vectors pET32-VP2 and pBV220-VP2 respectively. SDS-PAGE showed highlyefficient expression of codon-modified IBDV VP2 gene in E. coli. Western-blot analysis showed that the ex- pressed VP2 protein could react with the chicken against IBD serum. This study provided a basis for the studies of the structure and function of VP2 gene and IBD immune prevention and rapid diagnosis.

关 键 词：鸡传染性法氏囊病病毒 VP2 密码子优化 表达 

分 类 号：S852.659.4[农业科学—基础兽医学]