2.2.15细胞中HBV DNA前C和C区PCR扩增及其产物测序

AMPLIFICATION OF POLYMERASE CHAIN REACTION AND DIRECT SEQUENCING OF HEPATITS B VIRAL GENOME ENCOMPASSING PRE-C AND C REGION IN 2. 2. 15 CELLS

应用聚合酶链反应（PCR）及其扩增产物的直接测序技术对2．2．15细胞中乙型肝炎病毒（HBtV)基因前C和C区进行测序．结果表明，根据ayw亚型HBV基因设计的引物能够扩增2．215细胞中HBVDNA，扩增产物位于221－298碱基对之间，而对adr亚型HBVDNA无扩增作用，说明PCR扩增是特异性的，所测到的132个碱基序列与aywl亚型HBVDNA完全符合，提示22．15细胞中HBVDNA属aywl亚型．本文初步确定了22．15细胞中HBVDNA的亚型，对今后抗HBV药物（如反义寡核苷酸）在体外细胞培养中的研究具有重要的指导意义。

The sequence of hepatitis B viral genome in the 2. 2. 15 cells was determined amplification of polymerase chain reaction (PCR) and direct sequencing of its amplified products. Our resultsshowed that PCR amplification was specifical and HBV DNA in the 2. 2. 15 cells was from HBV withsurface antigen subtype aywl,because amplified products were located between bp 221 and bp 298 andthe sequence was in accord with subtype aywl. We first identifid the subtype of HBV genome in the2- 2. 15 cells, which might play an improtant role in researches on anti-HBV drugs in vitro cell culture assay system.