Identification and physical characterization of yeast maltase structural genes.
Identification and physical characterization of yeast maltase structural genes.
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酵母麦芽糖酶结构基因的鉴定和物理表征。
DOI：
10.1007/bf00425747
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Marmur,J
中科院分区：
文献类型：
--
作者：
Chow,T;Goldenthal,MJ;Cohen,JD;Hegde,M;Marmur,J
酵母基因组上至少5个未连接的MAL基因座（MAL 1至MAL 4和MAL 6）中的每一个都控制着合成可诱导的α-D-葡萄糖苷酶（麦芽糖酶）的能力。以MAL 6麦芽糖酶结构基因编码序列的一个亚克隆片段为探针，研究了不同链霉菌基因组中MAL结构基因家族的物理结构。Mal+菌株，每个携带一个遗传上确定的MAL基因座，与Mal-菌株杂交，并分析功能基因座和与该基因座上的麦芽糖酶结构基因互补的序列的分离行为。用BamH 1引物对减数分裂产物基因组DNA进行Southern印迹杂交，检测每个MAL位点的麦芽糖酶结构基因序列。这种限制性内切酶以前被证明在MAL 6基因座的范围外切割。这些实验的结果表明，每个MAL基因座包含至少一个与MAL 6同源的麦芽糖酶结构基因序列，缺乏功能性MAL基因座的酵母菌株可能含有或不含有相应的麦芽糖酶结构基因序列，MAL 1麦芽糖酶结构基因序列或其等位基因之一可以在所有实验室检测的酵母菌株中检测到，并且每个MAL基因座可以被鉴定为基因组DNA的特征性BamH 1片段，其包括麦芽糖酶结构基因，酵母菌株在它们携带的麦芽糖酶结构基因序列的数量上不同。通过使用本报告中描述的方法，可以鉴定对应于不同功能性MAL基因座并位于BamH 1产生的限制性片段内的那些。
Each of at least five unlinkedMALloci (MAL1throughMAL4andMAL6) on the yeast genome controls the ability to synthesize an inducible α-D-glucosidase (maltase). A subcloned fragment of the coding sequence of theMAL6maltase structural gene was used as a hybridization probe to investigate the physical structure of the family ofMALstructural genes in the genomes of differentSaccharomycesstrains. Mal+strains, each carrying a genetically definedMALlocus, were crossed with a Mal-strain and the segregation behavior of the functional locus and of sequences complementary to the maltase structural gene at that locus analyzed. The maltase structural gene sequences of eachMALlocus were detected by Southern blot hybridization using BamH1 digests of genomic DNA of the meiotic products. This restriction enzyme was previously shown to cleave outside the confines of theMAL6locus.The results of such experiments indicate that each MAL locus encompasses at least one maltase structural gene sequence homologous to that ofMAL6, that yeast strains that lack functionalMALloci may or may not contain the corresponding maltase structural gene sequence, that theMAL1maltase structural gene sequence or one of its alleles can be detected in all laboratory yeast strains examined and that eachMALlocus can be identified as a characteristic BamH1 fragment of genomic DNA which includes a maltase structural gene.Yeast strains vary in the number of maltase structural gene sequences that they carry. By using the approach described in this report, the ones corresponding to the different functionalMALloci and residing within a BamH1 generated restriction fragment can be identified.