Meyhack B, Bajwa W, Rudolph H, Hinnen A
EMBO J. 1982;1(6):675-80. doi: 10.1002/j.1460-2075.1982.tb01229.x.
We have cloned the structural genes for a regulated ( PHO5 ) and a constitutive ( PHO3 ) acid phosphatase from yeast by transformation and complementation of a yeast pho3 , pho5 double mutant. Both genes are located on a 5.1-kb BamHI fragment. The cloned genes were identified on the basis of genetic evidence and by hybrid selection of mRNA coupled with in vitro translation and immunoprecipitation. Subcloning of partial Sau3A digests and functional in vivo analysis by transformation together with DNA sequence analysis showed that the two genes are oriented in the order (5') PHO5 , PHO3 (3'). While the nucleotide sequences of the two coding regions are quite similar, the putative promoter regions show a lower degree of sequence homology. Partly divergent promoter sequences may explain the different regulation of the two genes.
我们通过酵母pho3、pho5双突变体的转化和互补,克隆了酵母中一个受调控的(PHO5)和一个组成型的(PHO3)酸性磷酸酶的结构基因。这两个基因都位于一个5.1kb的BamHI片段上。克隆的基因是根据遗传证据以及通过mRNA的杂交选择结合体外翻译和免疫沉淀来鉴定的。部分Sau3A消化产物的亚克隆以及通过转化进行的体内功能分析与DNA序列分析表明,这两个基因的排列顺序为(5')PHO5、PHO3(3')。虽然两个编码区的核苷酸序列非常相似,但推定的启动子区域显示出较低程度的序列同源性。部分不同的启动子序列可能解释了这两个基因的不同调控方式。