Barel I, Bignell G, Simpson A, MacDonald D
Department of Genetics, Cambridge, UK.
Curr Genet. 1988 Jun;13(6):487-94. doi: 10.1007/BF02427754.
From a gene bank of S. pombe DNA, a 5.6 kb clone was isolated which complemented mutants defective in glutamine synthetase (GS) activity. Sub-cloning fragments of this 5.6 kb clone showed that the complementing activity was localised in a 1.6 kb HindIII-AvaI fragment and a partial DNA sequence revealed an open reading frame preceded by TATA sequences and a TGACTA sequence. Plasmid constructs carrying up to 3.4 kb of DNA used to transform gln- strains gave transformants which showed a wide range of GS activity, in some cases 100 times the wild-type level. These constructs identify DNA sequences lying downstream from the putative coding sequence which have effects on the total amount of enzyme activity, but do not affect the control imposed by the nitrogen source on which the cells are grown.
从粟酒裂殖酵母DNA基因库中分离出一个5.6kb的克隆，它能互补谷氨酰胺合成酶（GS）活性缺陷的突变体。对该5.6kb克隆的亚克隆片段进行分析表明，互补活性定位于一个1.6kb的HindIII - AvaI片段上，部分DNA序列显示有一个前面带有TATA序列和TGACTA序列的开放阅读框。携带高达3.4kb DNA的质粒构建体用于转化gln-菌株，得到的转化体显示出广泛的GS活性范围，在某些情况下是野生型水平的100倍。这些构建体鉴定出位于推定编码序列下游的DNA序列，这些序列对酶活性的总量有影响，但不影响细胞生长所依赖的氮源对其施加的调控。
