Yao Y C, Lei Z H, Wang Z H, Li J X, Wang X M, Li H, Hu Y P
Department of Cell Biology, Second Military Medical University, Shanghai 200433, China.
Yi Chuan Xue Bao. 2000;27(11):941-6.
The 3'-truncated preS/S region from HBV genome, encoding a transcriptional transactivator, was cloned and a recombination expression vector for 3'-truncated HBV preS/S sequences under the control of the CMV promoter was constructed by recombination DNA techniques. Then, the expression vector DNA was microinjected into pronuclei of fertilized mouse oocytes. Founders of transgenic mice harbouring the recombination gene which can be expressed were screened by polymerase chain reaction (PCR) at genomic DNA level and confirmed by ELISA andlysis at protein level. Two of 15 mice in one series of microinjections showed the expression of 3'-truncated preS/S gene from HBV genome. The expression vector of 3'-truncated preS/S gene might be helpful for further studies of relationship between the expression production of 3'-truncated preS/S sequence and HBV-associated oncogenesis in vitro. The transgenic mice with expressing 3'-truncated preS/S gene will provide new insight into its roles in the development of human hepatocelluar carcinoma (HCC) in vivo.
克隆了来自乙肝病毒(HBV)基因组的3'端截短的前S/S区域,其编码一种转录反式激活因子,并通过重组DNA技术构建了一个在巨细胞病毒(CMV)启动子控制下的3'端截短的HBV前S/S序列的重组表达载体。然后,将表达载体DNA显微注射到受精小鼠卵母细胞的原核中。通过聚合酶链反应(PCR)在基因组DNA水平筛选携带可表达重组基因的转基因小鼠的奠基者,并通过酶联免疫吸附测定(ELISA)和蛋白质水平分析进行确认。在一系列显微注射的15只小鼠中,有2只显示出HBV基因组3'端截短的前S/S基因的表达。3'端截短的前S/S基因的表达载体可能有助于进一步研究3'端截短的前S/S序列的表达产物与体外HBV相关肿瘤发生之间的关系。表达3'端截短的前S/S基因的转基因小鼠将为其在体内人类肝细胞癌(HCC)发展中的作用提供新的见解。