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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Identification of two flavivirus-like genomes in the GB hepatitis agent.
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Identification of two flavivirus-like genomes in the GB hepatitis agent.

机译：GB肝炎病原中两个黄病毒样基因组的鉴定。

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摘要

A subtractive PCR methodology known as representational difference analysis was used to clone specific nucleotide sequences present in the infectious plasma from a tamarin infected with the GB hepatitis agent. Eleven unique clones were identified, seven of which were examined extensively. All seven clones appeared to be derived from sequences exogenous to the genomes of humans, tamarins, Saccharomyces cerevisiae, and Escherichia coli. In addition, sequences from these clones were not detected in plasma or liver tissue of tamarins prior to their inoculation with the GB agent. These sequences were detected by reverse transcription-PCR in acute-phase plasma of tamarins inoculated with the GB agent. Probes derived from two of the seven clones detected an RNA species of > or = 8.3 kb in the liver of a GB-agent-infected tamarin by Northern blot hybridization. Sequence analysis indicated that five of the seven clones encode polypeptides that possess limited amino acid identity with the nonstructural proteins of hepatitis C virus. Extension of the sequences found in the seven clones revealed that plasma from an infected tamarin contained two RNA molecules > 9 kb long. Limited sequence identity with various isolates of hepatitis C virus and the relative positions of putative RNA helicases and RNA-dependent RNA polymerases in the predicted protein products of these molecules suggested that the GB agent contains two unique flavivirus-like genomes.
机译：使用称为代表性差异分析的减法PCR方法从感染了GB肝炎病毒的猴克隆感染血浆中存在的特定核苷酸序列。鉴定出十一个独特的克隆，其中七个被广泛检查。所有七个克隆似乎都来自人类，猴，酿酒酵母和大肠杆菌的基因组的外源序列。另外，在用GB剂接种之前，在in猴的血浆或肝组织中未检测到来自这些克隆的序列。通过逆转录-PCR在接种GB试剂的agent猴急性期血浆中检测到这些序列。源自七个克隆中两个克隆的探针通过Northern blot杂交在被GB试剂感染的绢毛猴的肝脏中检测到大于或等于8.3 kb的RNA物种。序列分析表明，七个克隆中的五个编码的多肽与丙型肝炎病毒的非结构蛋白具有有限的氨基酸同一性。在七个克隆中发现的序列的扩展表明，来自被感染的绢毛猴的血浆中含有两个长于9 kb的RNA分子。与丙型肝炎病毒的各种分离物的序列同一性有限，以及在这些分子的预测蛋白产物中推定的RNA解旋酶和RNA依赖性RNA聚合酶的相对位置表明，GB剂包含两个独特的黄病毒样基因组。

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