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六种酿酒酵母菌株中第三条染色体的物理比较。

A physical comparison of chromosome III in six strains of Saccharomyces cerevisiae.

作者信息

Wicksteed B L, Collins I, Dershowitz A, Stateva L I, Green R P, Oliver S G, Brown A J, Newlon C S

机构信息

University of Aberdeen, Marischal College, U.K.

出版信息

Yeast. 1994 Jan;10(1):39-57. doi: 10.1002/yea.320100105.

Abstract

We have tested the clones used in the European Yeast Chromosome III Sequencing Programme for possible artefacts that might have been introduced during cloning or passage through Escherichia coli. Southern analysis was performed to compare the BamHI, EcoRI, HindIII and PstI restriction pattern for each clone with that of the corresponding locus on chromosome III in the parental yeast strain. In addition, further enzymes were used to compare the restriction maps of most clones with the map predicted by the nucleotide sequence (Oliver et al., 1992). Only four of 506 6-bp restriction sites predicted by the sequence were not observed experimentally. No significant cloning artefacts appear to disrupt the published sequence of chromosome III. The restriction patterns of six yeast strains have also been compared. In addition to two previously identified sites of Ty integration on chromosome III (Warmington et al., 1986; Stucka et al., 1989; Newlon et al., 1991), a new polymorphic site involving Ty retrotransposition (the Far Right-Arm transposition Hot-Spot, FRAHS) has been identified close to CRY1. On the basis of simple restriction polymorphisms, the strains S288C, AB972 and W303-1b are closely related, while XJ24-24a and J178 are more distant relatives of S288C. A polyploid distillery yeast is heterozygous for many polymorphisms, particularly on the right arm of the chromosome.

摘要

我们已经对欧洲酵母III号染色体测序计划中使用的克隆进行了检测,查找在克隆过程或通过大肠杆菌传代过程中可能引入的人为假象。进行了Southern分析,以比较每个克隆的BamHI、EcoRI、HindIII和PstI限制性酶切图谱与亲本酵母菌株III号染色体上相应位点的图谱。此外,还使用了其他酶来比较大多数克隆的限制性图谱与核苷酸序列预测的图谱(Oliver等人,1992年)。在序列预测的506个6碱基限制性位点中,只有4个未在实验中观察到。似乎没有明显的克隆假象扰乱已发表的III号染色体序列。还比较了6个酵母菌株的限制性图谱。除了先前在III号染色体上确定的两个Ty整合位点(Warmington等人,1986年;Stucka等人,1989年;Newlon等人,1991年)外,在CRY1附近还发现了一个涉及Ty逆转座的新多态性位点(最右侧臂转座热点,FRAHS)。基于简单的限制性多态性,菌株S288C、AB972和W303-1b亲缘关系密切,而XJ24-24a和J178是S288C较远的亲缘菌株。一种多倍体酿酒酵母在许多多态性方面是杂合的,特别是在染色体的右臂上。

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