Localization of yeast glucoamylase genes by PFGE and OFAGE.
Localization of yeast glucoamylase genes by PFGE and OFAGE.
复制标题
通过 PFGE 和 OFAGE 对酵母葡糖淀粉酶基因进行定位。
DOI：
10.1007/bf00405847
复制
发表时间：
1988
期刊：
影响因子：
2.5
通讯作者：
Marmur,J
中科院分区：
文献类型：
--
作者：
Pretorius,IS;Marmur,J
通过脉冲场凝胶电泳 (PFGE) 和同源场改变凝胶电泳 (OFAGE) 解析了两种密切相关的酵母菌株（淀粉分解糖化酵母和非淀粉分解酿酒酵母）的染色体。这两个菌株的电泳核型相同。已知染色体位置的 16 个克隆酵母基因用于通过 Southern 杂交分析来鉴定个体染色体。用S.的STA2葡糖淀粉酶基因的克隆片段重新检测Southern印迹。 diastaticus.STA2 与 STA1 和 STA3 以及来自两种酵母菌株的孢子形成特异性葡糖淀粉酶 (SGA) 基因表现出同源性。三个不连锁的同源基因 STA1 (DEX2,MAL5)、STA2 (DEX1) 和 STA3 (DEX3) 编码 S 中的胞外葡糖淀粉酶同工酶 GAI、GAII 和 GAIII。然后将 diastaticus 分别分配到 IV、II 和 XIV 号染色体。 SGA 基因，编码细胞内葡糖淀粉酶。糖化病和S. cerevisiae，被分配到 IX 号染色体。 STA 和 SGA 基因的电泳图谱是目前定位这些基因的唯一方法，因为葡糖淀粉酶阻遏基因（STA10、INH1 和/或 IST2）存在于大多数实验室菌株中。酿酒酵母和 SGA 表型仅在孢子形成期间可检测到。
Chromosomes of two closely related yeast strains, the amylolyticSaccharomyces diastaticusand the non-amylolyticSaccharomyces cerevisiae, were resolved by pulsed field gel electrophoresis (PFGE) and orthological field alteration gel electrophoresis (OFAGE). Electrophoretic karyotypes of these two strains are identical. Sixteen clonedSaccharomycesgenes of known chromosomal location were used to identify individual chromosomes by Southern hybridization analyses. The Southern blots were reprobed with a cloned fragment of theSTA2glucoamylase gene ofS. diastaticus.STA2 exhibits homology toSTA1 andSTA3 as well as the sporulation-specific glucoamylase (SGA) gene from bothSaccharomycesstrains. The three unlinked, homologous genes,STA1 (DEX2,MAL5),STA2 (DEX1) andSTA3 (DEX3) encoding the extracellular glucoamylase isozymes GAI, GAII and GAIII inS. diastaticuswere then assigned to chromosomes IV, II and XIV, respectively. The SGA gene, encoding an intracellular glucoamylase in bothS. diastaticusandS. cerevisiae, was assigned to chromosome IX. Electrophoretic mapping of the STA and SGA genes is at present the only way to localize these genes, since glucoamylase repressor gene(s) (STA10,INH1 and/orIST2) are present in most laboratory strains ofS. cerevisiaeand theSGAphenotype is only detectable during sporulation.