Chu M L, Myers J C, Bernard M P, Ding J F, Ramirez F
Nucleic Acids Res. 1982 Oct 11;10(19):5925-34. doi: 10.1093/nar/10.19.5925.
We report the cloning of five overlapping cDNAs bearing sequences specific for the human pro alpha 1(I) collagen chain. Poly-A RNA enriched for collagen sequences was purified from normal human fibroblasts and used as template to synthesize double stranded cDNA. The cDNA was inserted into the Eco RI site of pBR 322 by blunt-ending and dG:dC tailing. The clones were screened by colony hybridization using the original RNA population and the resulting five positive clones subjected to restriction endonuclease mapping analysis and DNA sequencing. These overlapping clones cover from residue 247 in the alpha chain to part of the 3' end untranslated region of the pro alpha 1(I) mRNA for a total of 3400 nucleotides.
我们报道了五条重叠互补DNA（cDNA）的克隆，这些cDNA带有人类原α1（I）胶原链特有的序列。从正常人成纤维细胞中纯化出富含胶原序列的聚腺苷酸RNA（Poly-A RNA），并将其用作模板来合成双链cDNA。通过平端连接和dG:dC加尾，将cDNA插入到pBR 322的Eco RI位点。使用原始RNA群体通过菌落杂交筛选克隆，并对得到的五个阳性克隆进行限制性内切酶图谱分析和DNA测序。这些重叠克隆覆盖了α链中第247位残基到原α1（I）mRNA 3'端非翻译区的一部分，总共3400个核苷酸。
