Cloning and characterization of five overlapping cDNAs specific for the human pro alpha 1(I) collagen chain.
Cloning and characterization of five overlapping cDNAs specific for the human pro alpha 1(I) collagen chain.
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人类原 α 1(I) 胶原链特异性的 5 个重叠 cDNA 的克隆和表征。
DOI：
10.1093/nar/10.19.5925
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发表时间：
1982
影响因子：
14.9
通讯作者：
F. Ramirez
中科院分区：
文献类型：
--
作者：
M. Chu;J. Myers;M. Bernard;J. Ding;F. Ramirez
我们报道了五个重叠的cDNA的克隆，这些cDNA含有人前α1(I)胶原链的特异性序列。从正常人成纤维细胞中提纯了富含胶原序列的Poly-A RNA，并以此为模板合成了双链cDNA。用末端钝化和DG：DC拖尾的方法将其插入到pBR 322的Eco RI位点。用原RNA群体进行菌落杂交筛选，得到5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从α链上的247个残基到原α1(I)基因3‘端非翻译区的一部分，总共有3400个核苷酸。
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.