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目的利用杆状病毒表达系统(BEVS)构建含乙型肝炎病毒核心抗原(HBcAg)基因的重组杆状病毒。方法采用PCR法从含乙型肝炎病毒全基因序列(adw)的质粒pHBV1中扩增出HBVC基因,亚克隆到pGEM-T载体,然后将HBVC基因插入载体pAcGHLT-A中,构建含有HBVC基因的重组质粒pAcGHLT-HBcAg,与苜蓿银纹夜蛾核型多角体病毒(AcNPV)联合转染秋粘虫细胞(Sf9),获得含有HBVC基因的重组杆状病毒。采用限制性内切酶酶切和DNA序列分析对重组病毒进行鉴定。结果利用杆状病毒表达系统,成功地构建了含HBVC基因的重组杆状病毒,感染昆虫细胞Sf9后,PCR检测可以扩增出相应大小的片段。结论利用BEVS成功构建了含有HBVC基因的重组杆状病毒,为进一步的研究奠定了基础。
Objective To construct a recombinant baculovirus containing hepatitis B virus core antigen (HBcAg) gene using baculovirus expression system (BEVS). Methods The HBVC gene was amplified by PCR from plasmid pHBV1 containing the full-length sequence of hepatitis B virus (adw), subcloned into pGEM-T vector, and inserted into vector pAcGHLT-A to construct a recombinant plasmid containing HBVC gene The recombinant plasmid pAcGHLT-HBcAg was co-transfected with Sf9 cells with Autographa californica nuclear polyhedrosis virus (AcNPV) to obtain recombinant baculovirus containing HBVC gene. Recombinant viruses were identified by restriction endonuclease digestion and DNA sequence analysis. Results The recombinant baculovirus containing HBVC gene was constructed successfully by baculovirus expression system. After infection with insect cell Sf9, PCR amplification could amplify the corresponding fragments. Conclusion The recombinant baculovirus containing HBVC gene was successfully constructed by BEVS, which laid the foundation for further research.