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利用酵母细菌穿梭载体构建粟酒裂殖酵母基因文库及其通过互补作用分离基因的应用。

Construction of a Schizosaccharomyces pombe gene bank in a yeast bacterial shuttle vector and its use to isolate genes by complementation.

作者信息

Beach D, Piper M, Nurse P

出版信息

Mol Gen Genet. 1982;187(2):326-9. doi: 10.1007/BF00331138.

Abstract

A gene bank of partial Sau3A restriction fragments of S. pombe DNA has been constructed in the plasmid vector, pDB248', which is capable of high frequency transformation of S. pombe. Procedures are described which enable plasmids to be recovered from S. pombe by their reintroduction into E. coli. These methods have been used to detect the S. pombe genes lys 1+, ade 6+ and his 2+ in the gene bank by complementation of mutant gene functions, and to physically isolate the lys 1+ gene.

摘要

利用质粒载体pDB248'构建了粟酒裂殖酵母DNA的部分Sau3A限制性片段基因文库,该载体能够高效转化粟酒裂殖酵母。文中描述了通过将质粒重新导入大肠杆菌从而从粟酒裂殖酵母中回收质粒的方法。这些方法已被用于通过突变基因功能互补在基因文库中检测粟酒裂殖酵母的lys 1 +、ade 6 +和his 2 +基因,并从物理上分离lys 1 +基因。

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