Surosky R T, Tye B K
Proc Natl Acad Sci U S A. 1985 Apr;82(7):2106-10. doi: 10.1073/pnas.82.7.2106.
We describe a simple method for the construction of large chromosomal deletions in yeast. Diploid yeast cells were transformed with DNA fragments that replace large regions of the chromosomes by homologous recombination. Using this method, we have constructed a telocentric chromosome III in which approximately equal to 100 kilobases (kb) of DNA has been removed from the left arm of the chromosome, so that the centromere is 12 kb from the left telomere. This telocentric chromosome is mitotically stable. Its rate of loss in a diploid strain is 2.5-7.4 X 10(-4) per cell division compared to a rate of loss of 0.36-1.8 X 10(-4) per cell division for a normal chromosome III. It also segregates 2+:2- with fidelity during meiosis. The construction of systematic deletions in a chromosome should be useful in determining the essential features for proper chromosomal segregation and replication.
我们描述了一种在酵母中构建大型染色体缺失的简单方法。用通过同源重组取代染色体大片段的DNA片段转化二倍体酵母细胞。使用这种方法，我们构建了一条端着丝粒染色体III，其中约100千碱基（kb）的DNA已从染色体左臂去除，使得着丝粒距左末端12 kb。这条端着丝粒染色体在有丝分裂中是稳定的。在二倍体菌株中，其每个细胞分裂的丢失率为2.5 - 7.4×10⁻⁴，而正常染色体III每个细胞分裂的丢失率为0.36 - 1.8×10⁻⁴。在减数分裂过程中，它也能以保真度以2+:2-的比例分离。在染色体中构建系统性缺失对于确定正确染色体分离和复制的基本特征应该是有用的。
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