通过重组筛选分离小鼠热休克基因（hsp68）。

Isolation of a mouse heat-shock gene (hsp68) by recombinational screening.

作者信息

Perry M D, Moran L A

出版信息

Gene. 1987;51(2-3):227-36. doi: 10.1016/0378-1119(87)90311-8.

Abstract

We have used cloned fragments from a Drosophila melanogaster hsp70 gene and a mouse hsp68 cDNA in recombinational screens of mouse genomic libraries. Using the mouse probe we have isolated two overlapping recombinant lambda phages comprising 22 kb of cloned DNA. Southern analysis has localized the homology with the Drosophila hsp70 coding region to a 2.2-kb fragment containing the mouse heat-shock gene. Insertion accompanying recombinational screening can disrupt interesting sequences; we have overcome this inconvenience by developing a simple one-step genetic selection for phage which have precisely excised the microplasmid probe.

摘要

我们利用果蝇热休克蛋白70（hsp70）基因的克隆片段和小鼠热休克蛋白68（hsp68）的互补DNA（cDNA）对小鼠基因组文库进行重组筛选。使用小鼠探针，我们分离出了两个重叠的重组λ噬菌体，它们包含22 kb的克隆DNA。Southern分析已将与果蝇hsp70编码区的同源性定位到一个包含小鼠热休克基因的2.2 kb片段上。重组筛选过程中的插入可能会破坏感兴趣的序列；我们通过开发一种简单的一步法遗传筛选方法克服了这一不便，该方法用于筛选已精确切除微质粒探针的噬菌体。