摘要
采用PCR亚克隆了山羊痘病毒P32基因片段,将其插入杆状病毒转座载体质粒pFast-BacⅠ中,构建了重组质粒pFastBac-P32;再将该质粒转化DH10 Bac感受态细菌,进行体内重组,然后经三重抗性及蓝白斑筛选,获得杆状病毒重组质粒Bacmid-P32;将Bacmid-P32转染Sf9细胞,获得重组杆状病毒;最后用SDS-PAGE和Western-blotting对P32基因的表达进行检测。结果表明,P32基因在重组杆状病毒中获得表达,且表达产物可以被多抗所识别。
The P32 gene of goatpox virus was amplified by PCR, and then cloned into a baculovirus expression vector pFastBac I . A recombinant plasmid pFastBac-P32 was constructed and then transformed into DH10Bac competent cells. White colonies of the recombinant Bacmid-P32 were obtained by screening with three antibiotics and selection of blue-white patches. After the Bacmid-P32 was transfected into Sf9 insect cells, the recombinant baculovirus was obtained by three passages. Finally, the expressed product from the P32 gene was detected by SDS-PAGE and Western-blotting. Result showed that the P32 gene was expressed successfully in
出处
《中国兽医科技》
CSCD
北大核心
2005年第12期950-953,共4页
Chinese Journal of Veterinary Science and Technology
关键词
山羊痘病毒
P32基因
杆状病毒
表达
goatpox the recombinant baculovirus. virus
P32 gene
baculovirus
expression
