中国株乙型肝炎病毒全基因组克隆的制备与鉴定

Construction and identification of HBV full-length genome clone of Chinese strain

目的:扩增全长HBV DNA基因组,建立中国株HBV感染性克隆,阐述HBV基因组变异对其不同基因生物学特性影响。方法:TD-PCR和XL-PCR技术一次扩增HBV全长基因组,克隆,PCR、酶切和测序鉴定。结果:通过PCR、酶切、测序鉴定和真核细胞转染分析,获得全长HBV基因组克隆。结论:获得了HBV全基因组克隆,在真核细胞中可以表达HBsAg,为建立HBV感染性克隆奠定物质基础。

Aim: To construct hepatitis B virus ( HBV) clone of Chinese strain and expatiate on how the variant of HBV genome influences biological characteristic of gene in HBV. Methods: The full-length HBV DNA was amplified with TD-PCR and XL-PCR, and then cloned them into Puc18 vector. Results: The full-length HBV clone after determination by enzyme digestion and sequence analysis was acguired. Conclusions:It showed that the expressed antigen has good antige-nisity and can give the basement for constructing the head to tail infectious HBV clone of Chinese strain.