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克隆的人类胎儿珠蛋白基因的分离与鉴定

Isolation and characterization of cloned human fetal globin genes.

作者信息

Ramirez F, Burns A L, Mears J G, Spence S, Starkman D, Bank A

出版信息

Nucleic Acids Res. 1979 Nov 10;7(5):1147-62. doi: 10.1093/nar/7.5.1147.

Abstract

Three clones containing both the human G gamma and A gamma globlin genes have been isolated and characterized from a library of DNA fragments generated by partial Eco RI digestion of cellular DNA using charon 4A phage as vector. Two of the clones (NY 2 and 3) are identical and have an insert of 14.0 kb. The third clone (NY 1) has a 15.4 kb insert by virtue of an extra 1.4 kb Eco RI fragment at its 5' most end. This clone also has a Kpn I site not present in the other two suggesting it is the product of the gamma gene on the opposite chromosome. Restriction analysis of the three clones indicates that the G gamma and A gamma genes are linked on a single continuous piece of DNA and are separated by 3.5 kb and each contains at least one large intervening sequence of 0.85 kg between the Bam HI and Eco RI sites. These findings in cloned DNA provide direct evidence for linkage and organization of the gamma genes in man.

摘要

已从以Charon 4A噬菌体为载体,通过细胞DNA的部分Eco RI消化产生的DNA片段文库中分离并鉴定出三个包含人类Gγ和Aγ球蛋白基因的克隆。其中两个克隆(NY 2和3)相同,插入片段为14.0 kb。第三个克隆(NY 1)在其最5'端有一个额外的1.4 kb Eco RI片段,因此插入片段为15.4 kb。该克隆还具有另外两个克隆中不存在的Kpn I位点,这表明它是来自相反染色体上γ基因的产物。对这三个克隆的限制性分析表明,Gγ和Aγ基因在一条连续的DNA片段上相连,间隔3.5 kb,并且在Bam HI和Eco RI位点之间各自至少包含一个0.85 kb的大间隔序列。克隆DNA中的这些发现为人类γ基因的连锁和组织提供了直接证据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a705/342293/abb6347dbdfe/nar00458-0049-a.jpg

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