随机扩增多态性DNA PCR在hiv -1感染者基因组分析中的应用

Felix O Aikhionbare , Cheryl Newman , Chad Womack , William Roth , Ketan Shah , Vincent C Bond
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引用次数: 2

摘要

随机扩增多态性DNA聚合酶链反应(RAPD-PCR)是一种用于检测基因组多态性的DNA指纹技术。我们使用16种不同的RAPD-PCR 10-mer引物扩增80例hiv -1感染者外周血单个核细胞(PBMC)的DNA。这些个体先前通过基因特异性引物的PCR鉴定为CCR5位点的杂合子(+/Δ32)和纯合子(+/+)。随机选择的16条RAPD引物中有4条在CCR5 (+/Δ32)杂合子和CCR5(+/+)纯合子之间产生了可区分的条带图谱。用四种RAPD引物中的一种对一些RAPD- pcr产物进行直接测序,得到了清晰可读的序列,但序列有限,与先前发表的CCR5等位基因(+/+)纯合子(98%相似)和(+/Δ32)杂合子(87%相似)的部分序列相似。因此,RAPD-PCR技术可用于鉴定可能与hiv -1感染易感性或hiv -1感染个体之间疾病进展差异相关的人类分子标记。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Application of random amplified polymorphic DNA PCR for genomic analysis of HIV-1-infected individuals

Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) is a DNA fingerprinting technique used to detect genomic polymorphisms. We employed sixteen different RAPD-PCR 10-mer primers to amplify DNA from the peripheral blood mononuclear cells (PBMC) of 80 HIV-1-infected individuals. These individuals were previously identified as either heterozygotes (+/Δ32) and homozygotes (+/+) for the CCR5 locus by PCR with gene specific primers. Four of the sixteen randomly selected RAPD primers produced distinguishable banding profiles between CCR5 (+/Δ32) heterozygotes and CCR5 (+/+) homozygotes. Direct sequencing of some RAPD-PCR products obtained with one of the four RAPD primers that were tested yielded clearly readable, but limited sequences, which were similar to portions of the previously published sequences for (+/+) homozygotes (98% similarity) and (+/Δ32) heterozygotes (87% similarity) of the CCR5 alleles. Thus, the RAPD-PCR technique may be useful for the identification of human molecular markers that may correlate with susceptibility to HIV-1-infection, or differences in disease progression among HIV-1-infected individuals.

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