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酿酒酵母质粒系统中,有丝分裂基因转换长度、共转换模式及相互重组发生率。

Mitotic gene conversion lengths, coconversion patterns, and the incidence of reciprocal recombination in a Saccharomyces cerevisiae plasmid system.

作者信息

Ahn B Y, Livingston D M

出版信息

Mol Cell Biol. 1986 Nov;6(11):3685-93. doi: 10.1128/mcb.6.11.3685-3693.1986.

Abstract

Plasmids capable of undergoing genetic exchange in mitotically dividing Saccharomyces cerevisiae cells were used to measure the length of gene conversion events, to determine patterns of coconversion when multiple markers were present, and to correlate the incidence of reciprocal recombination with the length of conversion tracts. To construct such plasmids, restriction site linkers were inserted both within the HIS3 gene and in the flanking sequences, and two different his3- alleles were placed in a vector. Characterization of the genetic exchanges in these plasmids showed that most occur with the conversion of one his3- allele. Many of these events included coconversions in which more than one marker along the allelic sequence was replaced. The frequency of coconversion decreased with the distance between two markers such that markers further than 1 kilobase apart were infrequently coconverted. From these results the average length of conversion was determined to be approximately 0.5 kilobase. Examination of coconversions involving three or more markers revealed an almost obligatory, simultaneous coconversion pattern of all markers. Thus, when two markers which flank an intervening marker are converted, the intervening marker is 20 times more likely to be converted than to remain unchanged. The results of these studies also showed that the incidence of reciprocal recombination, which accompanies more than 20% of the conversion events, is more frequent when the conversion tract is longer than average.

摘要

能够在有丝分裂的酿酒酵母细胞中进行基因交换的质粒被用于测量基因转换事件的长度,确定存在多个标记时的共转换模式,并将相互重组的发生率与转换片段的长度相关联。为构建此类质粒,在HIS3基因内部及其侧翼序列中插入了限制性酶切位点接头,并将两个不同的his3 - 等位基因置于载体中。对这些质粒中基因交换的表征表明,大多数交换发生在一个his3 - 等位基因的转换过程中。许多此类事件包括共转换,即等位基因序列上不止一个标记被替换。共转换的频率随着两个标记之间的距离增加而降低,以至于相距超过1千碱基的标记很少发生共转换。根据这些结果,确定转换的平均长度约为0.5千碱基。对涉及三个或更多标记的共转换的研究表明,所有标记几乎都呈现出一种必然的、同时共转换的模式。因此,当侧翼有一个中间标记的两个标记发生转换时,中间标记被转换的可能性比保持不变的可能性高20倍。这些研究结果还表明,超过20%的转换事件会伴随相互重组,当转换片段长于平均长度时,相互重组的发生率更高。

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