Dalgleish R, Trapnell B C, Crystal R G, Tolstoshev P
J Biol Chem. 1982 Nov 25;257(22):13816-22.
HpCl, a Charon 4A bacteriophage containing a 16.3-kilobase insert of human genomic DNA, has been identified as representing the 3' portion of the human alpha 2(I) collagen gene by hybrid selected translation and cross-reactivity with characterized sheep and chick collagen genes. Evaluation of HpCl by restriction endonuclease mapping and alpha 2(I) mRNA hybridization demonstrated that the alpha 2(I) mRNA coding regions are separated by noncoding regions including one of 2.7 kilobase pairs near the 3' end of the mRNA coding region. In addition, a 1.8-kilobase XbaI-BamHI fragment containing the 2' a 1.8-kilobase XbaI-BamHI fragment containing the 3' end of the alpha 2(I) mRNA coding region includes a region of DNA that is repeated many times throughout the human genome. Quantification of alpha 2(I) gene number, using the technique of DNA dot hybridization, with two EcoRI fragments from HpClo as probes, demonstrated that HpCl is represented once in the human genome. In this context, it is unlikely that human alpha 2(I) collagen chain production is modulated by differential transcription of multiple alpha 2(I) genes.
HpCl是一种Charon 4A噬菌体,含有一段16.3千碱基的人类基因组DNA插入片段,通过杂交选择翻译以及与已鉴定的绵羊和鸡胶原蛋白基因的交叉反应,已被确定代表人类α2(I)胶原蛋白基因的3'部分。通过限制性内切酶图谱分析和α2(I)mRNA杂交对HpCl进行评估表明,α2(I)mRNA编码区被非编码区隔开,包括在mRNA编码区3'端附近的一个2.7千碱基对的非编码区。此外,一个包含α2(I)mRNA编码区3'端的1.8千碱基XbaI - BamHI片段包含一个在整个人类基因组中多次重复的DNA区域。使用DNA点杂交技术,以来自HpClo的两个EcoRI片段作为探针,对α2(I)基因数量进行定量分析,结果表明HpCl在人类基因组中仅出现一次。在这种情况下,人类α2(I)胶原蛋白链的产生不太可能通过多个α2(I)基因的差异转录来调节。