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。
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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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VOLUME CONTENTS Comprehensive analysis of a large genomic sequence at the putative B-cell chronic lymphocytic leukaemia (B-CLL) tumour suppresser gene locus Mutational analysis within the 3′ region of the PKD1 gene in Japanese families Allelic polymorphisms in the transcriptional regulatory region of human SEL1L CUMULATIVE AUTHOR INDEX FOR MUTNOM 2000
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