多功能酵母高拷贝穿梭载体

Multifunctional yeast high-copy-number shuttle vectors.

作者信息

Christianson T W, Sikorski R S, Dante M, Shero J H, Hieter P

机构信息

Department of Microbiology, Southern Illinois University, Carbondale 62901.

出版信息

Gene. 1992 Jan 2;110(1):119-22. doi: 10.1016/0378-1119(92)90454-w.

DOI:10.1016/0378-1119(92)90454-w

PMID:1544568

Abstract

A set of four yeast shuttle vectors that incorporate sequences from the Saccharomyces cerevisiae 2 mu endogenous plasmid has been constructed. These yeast episomal plasmid (YEp)-type vectors (pRS420 series) differ only in their yeast selectable markers, HIS3, TRP1, LEU2 or URA3. The pRS420 plasmids are based on the backbone of a multifunctional phagemid, pBluescript II SK+, and share its useful properties for growth in Escherichia coli and manipulation in vitro. The pRS420 plasmids have a copy number of about 20 per cell, equivalent to that of YEp24. During non-selective yeast growth, pRS420 plasmids are lost through mitotic segregation at rates similar to other YEp vectors and yeast centromeric plasmid (YCp) vectors, in the range of 1.5-5% of progeny per doubling. The pRS420 series provides high-copy-number counterparts to the current pRS vectors [Sikorski and Hieter, Genetics 122 (1989) 19-27].

摘要

构建了一组四个酵母穿梭载体，它们整合了酿酒酵母2μm内源质粒的序列。这些酵母附加体质粒（YEp）型载体（pRS420系列）的区别仅在于其酵母选择标记，即HIS3、TRP1、LEU2或URA3。pRS420质粒基于多功能噬菌粒pBluescript II SK +的骨架构建，并具有其在大肠杆菌中生长和体外操作的有用特性。pRS420质粒每个细胞的拷贝数约为20个，与YEp24相当。在非选择性酵母生长过程中，pRS420质粒通过有丝分裂分离而丢失，其速率与其他YEp载体和酵母着丝粒质粒（YCp）载体相似，每代子代的丢失率在1.5 - 5%之间。pRS420系列为当前的pRS载体[Sikorski和Hieter，遗传学122（1989）19 - 27]提供了高拷贝数的对应物。