传染性法氏囊病病毒YL株VP2基因的克隆、序列分析及原核表达

Cloning,Sequencing and Prokaryotic Expression of VP2 Gene of Infectious Bursal Disease Virus YL Strain

克隆、序列分析及原核表达鸡传染性法氏囊病病毒(IBDV)VP2基因。根据GenBank已登录的IB-DV VP2基因序列,设计合成一对VP2基因特异性引物,应用RT-PCR技术从陕西省某鸡场分离的IBDV毒株中克隆VP2基因并进行序列分析。再将VP2基因亚克隆于原核表达载体pET-32a中,构建重组质粒pET-32a-VP2,经鉴定正确后转化大肠杆菌BL21菌株进行诱导表达,并进行SDS-PAGE检测。成功克隆IBDVYL毒株VP2全基因序列,核苷酸和氨基酸序列分析表明,YL株为IBDV超强毒株。经IPTG诱导后,SDS-PAGE分析,在宿主菌BL21中成功表达约为53 ku的VP2蛋白。IBDV VP2基因克隆表达成功,为IBDV的分子生物学特性研究提供资料,其表达产物为进一步制备抗IBDV单克隆抗体奠定了基础。

Cloning,sequencing and prokaryotic expression chicken infectious bursal disease virus（IBDV） VP2 gene were conducted.According to the published sequence of VP2 gene of IBDV,a pair of specific primers were designed and synthesized.The VP2 gene was amplified by RT-PCR from IBDV YL strain isolated from a Shaanxi chicken farm.VP2 gene sequence was analyzed and cloned into pET-32a vector.The prokaryotic expression plasmid pET-32a-VP2 containing VP2 gene was successfully constructed.The expression of recombinant plasmid pET-32-VP2 in E.coli BL21 was induced and detected by SDS-PACE analysis.VP2 gene of IBDV YL strain was successful cloned and sequenced.Nucleotide and amino acid sequence analysis showed that YL strain was a high virulent stain of IBDV.An expected protein 53 ku in size was expressed properly from E.coli BL21 by SDS-PAGE analysis.Successful cloning and expression of IBDV VP2 gene provided molecular biology characteristics.Furthermore,the VP2 protein can be used to prepare IBDV monoclonal antibody.