利用聚合酶链式反应（PCR）产物和酿酒酵母在体内生成破坏盒式结构。

Generation of disruption cassettes in vivo using a PCR product and Saccharomyces cerevisiae.

作者信息

Zaragoza Oscar

机构信息

Instituto de Investigaciones Biomédicas Alberto Sols CSIC-UAM, C/Arturo Duperier 4, Madrid 28029, Spain.

出版信息

J Microbiol Methods. 2003 Jan;52(1):141-5. doi: 10.1016/s0167-7012(02)00154-9.

DOI:10.1016/s0167-7012(02)00154-9

PMID:12401237

Abstract

A method to obtain disruption cassettes based on the homologous recombination in Saccharomyces cerevisiae is described. The disruption marker is amplified by PCR using oligonucleotides containing 50 bp homologous to the disruptable gene and 20 bp from the marker. The PCR product is cotransformed into yeast with a plasmid containing the gene. After recombination, a plasmid that carries the disruption cassette for the gene is produced.

摘要

描述了一种基于酿酒酵母同源重组获得破坏盒的方法。使用含有与可破坏基因同源的50 bp和来自标记的20 bp的寡核苷酸通过PCR扩增破坏标记。将PCR产物与含有该基因的质粒共转化到酵母中。重组后，产生携带该基因破坏盒的质粒。