Monomorphism in humans and sequence differences among higher primates for a sequence tagged site (STS) in homeo box cluster 2 as assayed by denaturing gradient electrophoresis.
Monomorphism in humans and sequence differences among higher primates for a sequence tagged site (STS) in homeo box cluster 2 as assayed by denaturing gradient electrophoresis.
复制标题
通过变性梯度电泳测定同源盒簇 2 中序列标记位点 (STS) 的人类单态性和高等灵长类动物之间的序列差异。
DOI：
10.1093/nar/18.5.1314
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发表时间：
1990
影响因子：
14.9
通讯作者：
Kidd,KK
中科院分区：
文献类型：
--
作者：
Ruano,G;Gray,MR;Miki,T;Ferguson-Smith,AC;Ruddle,FH;Kidd,KK
Denaturing Gradient Electrophoresis (DGE) to amplify defined primate HOX2 segments and to detect sequence differences among them. Ifpriming sites are conserved, homologous PCR products can be amplified from primate genomes via Cross-Species PCR (CS-PCR) using human primers [2]. When subject to DGE, sequence differences in the most labile domain of CSPCR products will change the gel position at which the molecules partially melt and stop migration [3]. We have sequenced a PstI fragment 4 kb upstream from HOX 2.2 and synthesized primers delimiting both halves of a 630 bp segment witiin it [4]. PCR on various unrelatedhumans and CSPCR on chimpanzee, gorilla, orangutan andgibbon yielded products of the same length for each primer pair (Fig. la, c). The distal half,'Pyg 3-2', is 300 bp long and the proximal,'Pyg 1-4', is 320 bp. The 630 bp segment and flanking primers (Pyg 1, 2 below) define a Sequence Tagged Site (STS)[5] for the HOX2 region on human chromosome 17 (17q21-q22). Nested oligonucleotides (Pyg 3, 4 below) can serve as probes for confirming theSTS.All products focus on DGE at 2.5-3.1 M urea, 14-18% fonnamide (Fig. lb, d). No polymorphism is detected in humans for either half. In Pyg3-2, gorilla melts at 1% denaturant less than human; chimpanzee, at 5% less (Fig. lb). Hence, with respect to human, chimpanzee difference (s) destabilize Pyg 3-2 more thangorilla difference (s). Orangutan and gibbon donot amplify, presumably due to mismatch at priming sites. For Pyg 1-4, gorilla and chimpanzee melt at about the same range as humans (Fig. ld). Orangutan and gibbon are least stable: orangutan melts at 1% denaturant less and gibbon at 5% less than the other primates.
