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在感染马传染性贫血病毒的马细胞中检测前病毒DNA。

Detection of proviral DNA in horse cells infected with equine infectious anemia virus.

作者信息

Rice N R, Simek S, Ryder O A, Coggins L

出版信息

J Virol. 1978 Jun;26(3):577-83. doi: 10.1128/JVI.26.3.577-583.1978.

Abstract

Equine infectious anemia virus (EIAV) recently has been shown to possess a high-molecular-weight RNA genome and a virion reverse transcriptase. We completed the demonstration that EIAV is a retrovirus by showing the presence of proviral DNA in equine cells infected in vitro, but not in normal horse DNA. These studies were performed by using a highly representative cDNA probe synthesized by the virion polymerase. It was found that this cDNA reassociated extensively, and with high thermal stability, with either viral RNA or DNA extracted from infected cells, but showed no detectable reassociation with DNA from uninfected horse cells. Similarly, sequences related to EIAV were neither found in the DNA of four other Equus species, nor in a variety of other mammals including sheep, cows, pigs- dogs, cats, and humans; nor did EIAV cDNA hybridize with a variety of other retrovirus RNAs. These experiments were performed under conditions of very low stringency to enable detection of distantly related sequences, with a sufficient ratio of DAN to cDNA to allow detection of less than one viral copy per haploid genome. We conclude that EIAV is not an endogenous virus of the horse or of the other species tested.

摘要

马传染性贫血病毒(EIAV)最近被证明拥有一个高分子量的RNA基因组和一种病毒粒子逆转录酶。通过显示在体外感染的马细胞中存在前病毒DNA,而在正常马的DNA中不存在,我们完成了EIAV是一种逆转录病毒的证明。这些研究是通过使用由病毒粒子聚合酶合成的高度代表性的cDNA探针进行的。发现这种cDNA与从感染细胞中提取的病毒RNA或DNA广泛重新结合,并且具有高热稳定性,但与未感染马细胞的DNA没有可检测到的重新结合。同样,在其他四种马属动物的DNA中,以及在包括绵羊、牛、猪、狗、猫和人类在内的各种其他哺乳动物的DNA中,均未发现与EIAV相关的序列;EIAV cDNA也未与各种其他逆转录病毒RNA杂交。这些实验是在非常低的严谨性条件下进行的,以能够检测远缘相关序列,DNA与cDNA的比例足够高,以允许检测每个单倍体基因组中少于一个病毒拷贝。我们得出结论,EIAV不是马或所测试的其他物种的内源性病毒。

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