Synthesis and analysis of a 640-bp pol region of novel human endogenous retroviral sequences and their evolutionary relationships.
Synthesis and analysis of a 640-bp pol region of novel human endogenous retroviral sequences and their evolutionary relationships.
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新型人内源性逆转录病毒序列的 640 bp pol 区域的合成和分析及其进化关系。
DOI：
10.1006/viro.1996.0087
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Faras,AJ
中科院分区：
文献类型：
--
作者：
Li,MD;Lemke,TD;Bronson,DL;Faras,AJ
对47个逆转录元件进行的核苷酸序列比较发现了两个非常保守的区域，它们之间的跨度约为640个碱基对，这两个区域以前没有报道过。构建了一组与这两个保守区同源的5‘-MOP-2和3’-MOP-2混合寡核苷酸引物，用于反转录元素的检测。用MOPS-2进行PCR扩增时，人、小鼠基因组DNA和HIV-1前病毒DNA扩增出约0.64kb的产物，而阴性对照质粒DNA扩增不出产物。从人LUC-1畸胎癌细胞DNA中扩增出mOPS-2扩增产物，并进行亚克隆和测序。5个克隆的大小约为0.64kb，与GenBank中所有条目的序列比较表明，这5个克隆与人内源性逆转录病毒K(Herv-K)家族的HERV-K10和HM-16的相应区域有最高的同源性，在64.31~98.65%之间。经过严格的Southern杂交证明，这五个克隆在所有测试的人类DNA中都存在。通过系统发育分析，将3个克隆归入Herv-K家族，将2个克隆归入一个新的人类内源性逆转录元件家族，确定了这些克隆与其他逆转录元件等效区的进化关系。此外，克隆的Herv-(K)73含有较小的PV1bpol片段，据报道，该片段选择性地在真性红细胞增多症患者的血白细胞中表达。
Nucleotide sequence comparisons of thepolgene among 47 retroelements identified two very conserved regions, separated by a span of approximately 640 bp, that have not been previously reported. A set of mixed oligonucleotide primers, 5′-MOP-2 and 3′-MOP-2, homologous to these two conservedpolregions was constructed for use in detection of retroelements. When MOPs-2 were employed in PCR amplification studies, products of about 0.64 kb in size were amplified from human and mouse genomic DNAs and from HIV-1 proviral DNA, but not from negative control plasmid DNAs. The PCR products amplified with MOPs-2 from human LuC-1 teratocarcinoma cell DNA were subcloned and sequenced. Five clones of approximately 0.64 kb in size were identified, and sequence comparisons with all entries in GenBank indicated that these five clones have highest homology, in a range of 64.31 to 98.65%, with the correspondingpolregion of HERV-K10 and HM-16 of the human endogenous retrovirus-K (HERV-K) family. Southern hybridizations at high stringency demonstrated that these five clones are present in all human DNAs tested. The evolutionary relationships of these clones with the equivalentpolregion of other retroelements were defined by phylogenetic analyses that placed three clones into the HERV-K family and two clones into a new family of human endogenous retroelements. In addition, clone HERV-(K)73 contains the smaller PV1bpolsegment that was reported to be selectively expressed in blood leukocytes of patients with polycythemia vera.