THE DUCK HEPATITIS B-VIRUS P-GENE CODES FOR PROTEIN STRONGLY ASSOCIATED WITH THE 5'-END OF THE VIRAL-DNA MINUS STRAND
THE DUCK HEPATITIS B-VIRUS P-GENE CODES FOR PROTEIN STRONGLY ASSOCIATED WITH THE 5'-END OF THE VIRAL-DNA MINUS STRAND
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DOI：
10.1016/0042-6822(88)90518-1
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发表时间：
1988-10-01
期刊：
影响因子：
3.7
通讯作者：
SCHALLER, H
中科院分区：
文献类型：
--
作者：
BOSCH, V;BARTENSCHLAGER, R;SCHALLER, H
针对鸭B肝炎病毒（DHBV）P基因翻译产物的不同片段引发的许多抗血清用于免疫沉淀与DHBV DNA负链的5“-末端共价结合的蛋白。为了监测的目的，携带这种蛋白质的小DNA负链片段被放射性标记。所有的P特异性抗血清特异性免疫沉淀该DNA片段，证明附着于免疫沉淀的DNA片段的蛋白质种类是DHBV P基因的产物。DNA负链片段-蛋白质复合物在SDS凝胶中的电泳行为表明，它主要以聚集体的形式存在。然而，一小部分由携带P基因蛋白的DNA负链片段组成，仅在P基因的5“区域内编码。这表明，不同的P基因蛋白，可能共价结合在一个共同的区域，随后加工，结合到5“-端的DHBV DNA负链。DHBV P基因可能编码与病毒相关的逆转录酶和DNA聚合酶活性。使用P基因特异性抗血清，不可能检测到游离形式的推定的P基因编码的聚合酶蛋白，即，不与病毒DNA结合这可能是由于灵敏度不足或聚合酶蛋白是异质的和/或聚集的。此外，基因组结合蛋白本身可能具有聚合酶活性。
A number of antisera, elicited against different segments of the duck hepatitis B virus (DHBV) P-gene translation product, were used to immunoprecipitate the protein that is covalently bound to the 5''-end of the DHBV DNA minus strand. For monitoring purposes, a small DNA minus-strand fragment, carrying this protein, was radioactively labeled. All of the P-specific antisera specifically immunoprecipitated this DNA fragment demonstrating that the protein species attached to the immunoprecipated DNA-fragment were products of the DHBV P-gene. The electrophoretic behavior, in SDS gels, of the DNA minus-strand fragment-protein complex indicated that it was present mostly in the form of aggregates. However, a small fraction consisted of DNA minus-strand fragments carrying P-gene proteins, encoded solely within the 5''-region of the P-gene. This indicated that different P-gene proteins, presumably covalently bound at a common region and subsequently processed, were bound to the 5''-end of the DHBV DNA minus strand. The DHBV P-gene presumbaly codes for the virus-associated reverse transcriptase and DNA polymerase activities. Using the P-gene-specific antisera, it was not possible to detect putative P-gene-coded polymerase proteins in a free form, i.e., not bound to viral DNA. This may be due to the insufficient sensitivity or to the polymerase protein(s) being heterogeneous and/or aggregated. In addition, it is possible that the genome-bound protein itself may have polymerase activity.