HBV前C/C基因EB病毒载体的构建及在HepG2细胞中的表达

目的研究ＷＢＶ前Ｃ／Ｃ区基因变异的生物学意义。方法构建ＨＢＶ前Ｃ／Ｃ区基因ＥＢ病毒表达载体，采用脂质体介导方法将重组质粒转染到ＨｅｐＧ２细胞，并表达ＨＢｅＡｇ。结果重组质粒经ＰＣＲ和酶切鉴定均阳性，转染的细胞目的ＤＮＡ和表达蛋白检测均阳性。结论采用ＥＢ病毒载体构建ＨＢＶ前Ｃ／Ｃ基因的表达载体，能在ＨｅｐＧＺ细胞中稳定表达目的蛋白。由于ＥＢ病毒载体以附着体的形式复制，不整合于宿主染色体基因中，拷贝数稳定，适合于体外研究ＨＢＶＣ／Ｃ基因变异的生物学意义。

Construction of Esptein-Barr viral vector with hepatitis B virus pre C/C gene and its expression in HepG2 cells

Objective To study the biological significance of HBV preC/C gene mutation. Methods EB viral expression vector withHBV preC/C gene was constructed and transfected into HepG2 cells by means of lipoinfectin induction, and HBeAg was expressedin the transfected cells. Results The recombinant plasmid was detected positive by means of PCR and enzyme digestion, the goalDNA was contained and the goal protein was expressed in the transfected cells. Conclusion EB viral expression vector with HBVpreC/C gene can express the goal protein in HepG2 cells steadily. Because GB viral vector do not integrated into the hostchrolnogene and its copies keep steady. it can be used to study the biological signitlcance of HBV preC/C gene mutation in vitro.