在酵母重组缺陷型菌株中构建小鼠酵母人工染色体文库。

Construction of a mouse yeast artificial chromosome library in a recombination-deficient strain of yeast.

作者信息

Chartier F L, Keer J T, Sutcliffe M J, Henriques D A, Mileham P, Brown S D

机构信息

Department of Biochemistry and Molecular Genetics, St Mary's Hospital Medical School, London, UK.

出版信息

Nat Genet. 1992 May;1(2):132-6. doi: 10.1038/ng0592-132.

DOI:10.1038/ng0592-132

PMID:1302006

Abstract

We have constructed a new generation yeast artificial chromosome (YAC) library from female C57BL/10 mice in a recombination-deficient strain of Saccharomyces cerevisiae carrying a mutation in the RAD52 gene. The YAC library contains 41,568 clones with an average insert size of 240 kilobases, representing a greater than threefold coverage of the mouse genome. Currently, the library can be screened by polymerase chain reaction and we have isolated positive clones at a number of loci in the mouse genome. This rad52 library should enable a long-term assessment of the effect of one of the yeast recombination pathway genes on both, genome-wide YAC clone stability and the frequency of chimaeric clones.

摘要

我们在携带RAD52基因突变的酿酒酵母重组缺陷菌株中，构建了来自雌性C57BL/10小鼠的新一代酵母人工染色体（YAC）文库。该YAC文库包含41,568个克隆，平均插入片段大小为240千碱基，代表了小鼠基因组三倍以上的覆盖率。目前，该文库可通过聚合酶链反应进行筛选，我们已经在小鼠基因组的多个位点分离出了阳性克隆。这个rad52文库应该能够长期评估酵母重组途径基因之一对全基因组YAC克隆稳定性和嵌合克隆频率的影响。