乙型肝炎病毒3’末端缺失的preS/S基因在转基因小鼠中的表达

Expression of 3'-truncated preS/ S Gene in Transgenic Mice

从乙型肝炎病毒 ａｄｒ亚型的基因组 ＤＮＡ中分离 ３’末端缺失的ｐｒｅＳ／Ｓ基因的 ＤＮＡ片段，构建了由ＣＭＶ启动子控制的真核表达载体。采用受精卵显微注射方法，获得了基因组整合有３’末端缺失的ｐｒｅＳ／Ｓ基因的２个转基因小鼠品系。在不同的时间点采取血清进行了ＥＬＩＳＡ分析，发现在这２个小鼠品系中３’末端缺失的ｐｒｅＳ／Ｓ基因可被表达，而且呈稳定状态。此小鼠品系的建立，对于探讨乙型肝炎病毒３’末端缺失的ｐｒｅＳ／Ｓ基因的表达产物在体内的生物学作用及其与肝细胞内细胞癌基因的转录激活之间的关系有重要意义。

The 3'-truncated preS/S region from HBV genome, encoding a trans criptional 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 preduction of 3'-truncared preS/ S sequence and HBV-associated onco- genesis 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.