通过酵母中的转化相关重组直接分离人类BRCA2基因。
Direct isolation of human BRCA2 gene by transformation-associated recombination in yeast.
作者信息
Larionov V, Kouprina N, Solomon G, Barrett J C, Resnick M A
机构信息
Laboratories of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
出版信息
Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7384-7. doi: 10.1073/pnas.94.14.7384.
Abstract
Mutant forms of the BRCA2 gene contribute significantly to hereditary breast cancer. Isolation of the normal and mutant forms of the BRCA2 gene with its natural promoter would greatly facilitate analysis of the gene and its contribution to breast cancer. We have accomplished the direct isolation of the 90-kb gene from total human DNA by transformation-associated recombination in yeast using a small amount of 5' and 3' BRCA2 sequence information. Because the entire isolation procedure of a single chromosomal gene could be accomplished in approximately 2 weeks, the transformation-associated recombination cloning approach is readily applicable to studies of chromosome alterations and human genetic diseases.
摘要
BRCA2基因的突变形式在遗传性乳腺癌中起重要作用。分离具有天然启动子的BRCA2基因的正常和突变形式将极大地促进对该基因及其对乳腺癌作用的分析。我们利用少量5'和3' BRCA2序列信息，通过酵母中的转化相关重组从人总DNA中直接分离出了90 kb的基因。由于单个染色体基因的整个分离过程可在约2周内完成，因此转化相关重组克隆方法很容易应用于染色体改变和人类遗传疾病的研究。
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