Chow T, Goldenthal M J, Cohen J D, Hegde M, Marmur J
Mol Gen Genet. 1983;191(3):366-71. doi: 10.1007/BF00425747.
Each of at least five unlinked MAL loci (MAL1 through MAL4 and MAL6) on the yeast genome controls the ability to synthesize an inducible alpha-D-glucosidase (maltase). A subcloned fragment of the coding sequence of the MAL6 maltase structural gene was used as a hybridization probe to investigate the physical structure of the family of MAL structural genes in the genomes of different Saccharomyces strains. MAL+ strains, each carrying a genetically defined MAL locus, 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 each MaL locus 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 the MAL 6 locus. The results of such experiments indicate that each MAL locus encompasses at least one maltase structural gene sequence homologous to that of MAL6, that yeast strains that lack functional MAL loci may or may not contain the corresponding maltase structural gene sequence, that the MAL1 maltase structural gene sequence or one of its alleles can be detected in all laboratory yeast strains examined and that each MAL locus 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 functional MAL loci and residing within a BamH1 generated restriction fragment can be identified.
酵母基因组上至少五个不连锁的MAL位点(MAL1至MAL4和MAL6)中的每一个都控制着诱导性α-D-葡萄糖苷酶(麦芽糖酶)的合成能力。MAL6麦芽糖酶结构基因编码序列的一个亚克隆片段被用作杂交探针,以研究不同酿酒酵母菌株基因组中MAL结构基因家族的物理结构。将每个携带一个遗传定义的MAL位点的MAL+菌株与一个MAL-菌株杂交,并分析功能位点以及该位点与麦芽糖酶结构基因互补的序列的分离行为。通过使用减数分裂产物基因组DNA的BamH1酶切片段进行Southern印迹杂交,检测每个MAL位点的麦芽糖酶结构基因序列。先前已证明这种限制酶在MAL 6位点范围之外切割。此类实验结果表明,每个MAL位点都包含至少一个与MAL6同源的麦芽糖酶结构基因序列,缺乏功能性MAL位点的酵母菌株可能含有也可能不含有相应的麦芽糖酶结构基因序列,在所检测的所有实验室酵母菌株中都能检测到MAL1麦芽糖酶结构基因序列或其等位基因之一,并且每个MAL位点都可被鉴定为基因组DNA的一个特征性BamH1片段,其中包括一个麦芽糖酶结构基因。酵母菌株所携带的麦芽糖酶结构基因序列数量各不相同。通过使用本报告中描述的方法,可以鉴定出与不同功能性MAL位点相对应且位于BamH1产生的限制片段内的那些序列。