DIRECTION OF CHROMOSOME REARRANGEMENTS IN SACCHAROMYCES-CEREVISIAE BY USE OF HIS3 RECOMBINATIONAL SUBSTRATES
DIRECTION OF CHROMOSOME REARRANGEMENTS IN SACCHAROMYCES-CEREVISIAE BY USE OF HIS3 RECOMBINATIONAL SUBSTRATES
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DOI：
10.1128/mcb.8.10.4370
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发表时间：
1988-10-01
影响因子：
5.3
通讯作者：
DAVIS, RW
中科院分区：
文献类型：
--
作者：
FASULLO, MT;DAVIS, RW
我们使用的his3重组基板（his3片段）直接大染色体间（易位）和染色体内（缺失和串联重复）在酵母酿酒酵母重排。在野生型HIS3基因完全缺失的菌株中，首先通过同源重组将his3片段（一个含有5 ″氨基酸编码序列的缺失，另一个含有3 ″氨基酸编码序列的缺失）置于预选位点。选择通过两个his3片段之间的自发有丝分裂重组产生的His+回复突变体。该策略用于指导RAD 52+和rad52突变株中的重排。易位发生在RAD52+遗传背景中，并通过酵母染色体DNA的正交场交替凝胶电泳和标准遗传技术进行表征。还鉴定了一种意外的易位，其中HIS3序列扩增。GAL（7，10，1）位点的两种类型的串联重复也被定向，和一种类型的未观察到在rad52突变体。解释这些差异的机制进行了讨论。
We used the his3 recombinational substrates (his3 fragments) to direct large interchromosomal (translocations) and intrachromosomal (deletions and tandem duplications) rearrangements in the yeast Saccharomyces cerevisiae. In strains completely deleted for the wild-type HIS3 gne, his3 fragments, one containing a deletion of 5'' amino acid coding sequences and the other containing a deletion of 3'' amino acid coding sequences, were first placed at preselected sites by homologous recombination. His+ revertants that arose via spontaneous mitotic recombination between the two his3 fragments were selected. This strategy was used to direct rearrangements in both RAD52+ and rad52 mutant strains. Translocations occurred in the RAD52+ genetic background and were characterized by orthogonal field alternating gel electrophoresis of yeast chromosomal DNA and by standard genetic techniques. An unexpected translocation was also identified in which HIS3 sequences were amplified. Two types of tandem duplications of the GAL(7, 10, 1) locus were also directed, and one type was not observed in rad52 mutants. Recombination mechanisms are discussed to account for these differences.