Siddiqui A, Jameel S, Mapoles J
Proc Natl Acad Sci U S A. 1986 Feb;83(3):566-70. doi: 10.1073/pnas.83.3.566.
A series of recombinant plasmid vectors containing hepatitis B virus (HBV) DNA sequences was constructed to study the biosynthesis of the hepatitis B virus surface antigen (HBsAg) RNA and to locate transcriptional control elements involved in the regulation of the S and pre-S DNA sequences. We examined the transcription of the HBsAg gene in permanent cell lines that were developed by transfecting with recombinant vectors containing HBV sequences and the neomycin gene followed by G418 selection. We further defined the promoter activities upstream of and within the pre-S sequences using the assayable chloramphenicol acetyltransferase gene. Results obtained from S1 nuclease digestion and primer extension suggest that HBsAg transcripts are initiated at multiple sites in the pre-S region and from a site upstream of the pre-S region. Chloramphenicol acetyltransferase assays indicate that DNA sequences within and upstream of the pre-S region contain promoter activities and that the "TATA" sequence-containing promoter and the internal promoter show similar levels of activities in CV-1 cells and several other cell lines tested.
构建了一系列含有乙型肝炎病毒（HBV）DNA序列的重组质粒载体，以研究乙型肝炎病毒表面抗原（HBsAg）RNA的生物合成，并定位参与S和前S DNA序列调控的转录控制元件。我们检测了通过转染含有HBV序列和新霉素基因的重组载体，随后进行G418筛选而建立的永久细胞系中HBsAg基因的转录情况。我们使用可检测的氯霉素乙酰转移酶基因进一步确定了前S序列上游和内部的启动子活性。S1核酸酶消化和引物延伸实验结果表明，HBsAg转录本在前S区域的多个位点以及前S区域上游的一个位点起始。氯霉素乙酰转移酶分析表明，前S区域内部和上游的DNA序列具有启动子活性，并且含有“TATA”序列的启动子和内部启动子在CV-1细胞及其他几种测试细胞系中表现出相似的活性水平。
