Molecular cloning and carboxyl-propeptide analysis of human type III procollagen.
Molecular cloning and carboxyl-propeptide analysis of human type III procollagen.
复制标题
人 III 型前胶原的分子克隆和羧基前肽分析。
DOI：
10.1093/nar/12.24.9383
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发表时间：
1984
影响因子：
14.9
通讯作者：
Myers,JC
中科院分区：
文献类型：
--
作者：
Loidl,HR;Brinker,JM;May,M;Pihlajaniemi,T;Morrow,S;Rosenbloom,J;Myers,JC
在交叉杂交条件下筛选由正常成纤维细胞(GM3348)和横纹肌肉瘤(CCL136)mRNA制备的两个人cDNA文库，与编码禽类III型前胶原COOH前肽的外显子2和3的基因组片段(Yamada，Y.，Mudryj，M.，Sullivan，M.和deCrombrugghe，B.(1983)J.Biol.化学 258, 2758–2761）。从 CCL136 文库中分离出一个含有 1.12 kb 插入片段的 cDNA 克隆，随后用于鉴定具有 2.4 kb 插入片段的 GM3348 衍生克隆。人类和鸟类 III 型 C 端前肽的比较显示，三螺旋区末端半胱氨酸后面的前 54 个氨基酸的差异比这些物种的 α1(I) 和 α2(I) 前胶原链中存在的差异大得多。对正常人成纤维细胞和肿瘤细胞系的聚 (A+) RNA 的分析表明，它们的 α1(I)、α2(I) 和 α1(III) mRNA 的相对量存在很大差异。此外，正如我们之前报道的 α1(I) 和 α2(I) 转录本，多个 mRNA 也与克隆的 α1(III) DNA 杂交。
Two human cDNA libraries prepared from normal fibroblast (GM3348) and rhabdomyosarcoma (CCL136) mRNAs were screened under cross hybridization conditions with a genomic fragment coding for exons 2 and 3 of the avian type III procollagen COOH-propeptide (Yamada, Y., Mudryj, M., Sullivan, M. and deCrombrugghe, B. (1983) J. Biol . Chem. 258, 2758–2761). One cDNA clone containing a 1.12 kb insert was isolated from the CCL136 library and later used to identify a GM3348 derived clone with a 2.4 kb insert. Comparison of the human and avian type III C-terminal propeptides revealed much more divergence in the first 54 amino acids following the terminal cysteine of the triple helical region than is present in the α1(I) and α2(I) procollagen chains of these species. Analysis of poly (A+) RNA from normal human fibroblast and tumor cell lines showed that they differed greatly in the relative amounts of α1(I), α2(I), and α1(III) mRNAs. Furthermore, as we previously reported for the α1(I) and α2(I) transcripts, multiple mRNAs also hybridize to the cloned α1(III) DNA.