摘要
根据GenBank上登录的Ⅰ型鸭肝炎病毒(DHV-Ⅰ)基因组序列,设计一对特异性引物。通过RT-PCR的方法扩增DHV-Ⅰ(A66株)VP3基因并将其5′端起始处稀有密码子同义突变。用限制性内切酶BamHⅠ/SalⅠ消化VP3基因片段和表达载体pGEX-6p-1后构建重组表达质粒pGEX-VP3,转化E.coliBL21(DE3)。经IPTG诱导后,SDS-PAGE分析表明,VP3基因在大肠埃希菌中大量表达,表达产物的分子质量约为52 ku。Western blot检测表明,表达产物能与DHV-Ⅰ阳性血清发生反应,具有反应原性。
A pair of primers were designed and synthesized according to duck hepatitis virus 1 genome sequence in GenBank. VP3 gene of DHV- Ⅰ of A66 strain was amplified by RT-PCR and the mutation of the rare codon exited in 5′region was compeleted by the method of PCR. The vPa gene is subcoloned into the vector of pGEX-6p-1. The recombiant plasmid was transformed into the E. coli BL21 （DE3） component cells. SDS-PAGE analysis showed that the molecular weight of fusion protein GST-VP3 was 52 ku with highly expressed in E. coli BL21（DE3）. The protein can be recognized by antisera of DHV-Ⅰ , indicating that the fusion protein had good reactinogenicity.
出处
《动物医学进展》
CSCD
北大核心
2009年第7期49-52,共4页
Progress In Veterinary Medicine
