利用神经孢子虫孢子杀手菌株获得着丝粒连锁数据而无需解剖腹膜。

D D Perkins, N B Raju, V C Pollard, J L Campbell, A M Richman
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引用次数: 12

摘要

使用着丝粒连接的孢子杀手基因Sk可以大大减少获得四分体数据的工作量,否则需要有序地解剖完整的线性八孢子囊。杂合杂交用于孢子杀手(SkK X SkS)和待测标记。在这样的杂交中,只有SkK子囊孢子存活。每个子囊的4个活子囊孢子(SkK)和4个流产子囊孢子(SkS)作为一个物理上紊乱的群体从胚周射出。对单个子囊的4个存活的SkK子囊孢子进行萌发和评分。SkK在第一次减数分裂时从SkK中分离出来。如果两个标记等位基因都存在于存活的产物中,那么它们一定是在第二次分裂时彼此分离的。四孢子(Fsp)基因被用来消除一个减数分裂后的核分裂,因此每个子囊只需要对两个子囊孢子进行评分。孢子杀手方法在绘制着丝粒区域紧密连接的基因图谱、识别染色体臂上较远的基因、获得减数分裂交叉的信息以及比较不同物种间的联系方面非常有用。
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Use of Neurospora spore killer strains to obtain centromere linkage data without dissecting asci.

Use of a centromere-linked Spore killer gene Sk reduces manyfold the labor involved in obtaining tetrad data that would otherwise require ordered dissection of intact linear eight-spored asci. Heterozygous crosses are made for Spore killer (SkK X SkS) and for markers to be tested. In such crosses only SkK ascospores survive. The four viable (SkK) and four aborted (SkS) ascospores of each ascus are ejected from the perithecium as a physically disordered group. The four surviving SkK ascospores of individual asci are germinated and scored. SkK segregates from SkS at the first meiotic division. If both marker alleles are represented in the surviving products, they must therefore have segregated from one another at the second division. Four-spore (Fsp) genes have been used to eliminate one postmeiotic nuclear division, so that only two ascospores per ascus need to be scored. The Spore killer method has been useful for mapping closely linked genes in centromere regions, for identifying genes that are far out on chromosome arms, for obtaining information on meiotic crossing-over, and for comparing linkages in different species.

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Chromosome segregation from cell hybrids. II. Do differences in parental cell growth rates and phase times determine direction of loss? Use of Neurospora spore killer strains to obtain centromere linkage data without dissecting asci. Ascus development in two temperature-sensitive four-spore mutants of Neurospora crassa. Murine plasmacytomas constitute a class of natural heat-shock variants in which the major inducible hsp-68 gene is not expressed. Role of heterochromatin during preferential 9q;22q translocation in chronic myelogenous leukemia.
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