Ishibashi O, Shishido K
Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan.
Curr Genet. 1994 Jan;25(1):30-3. doi: 10.1007/BF00712963.
It was shown by a plasmid exchange procedure that the Ras-encoding cDNA of the basidiomycete Lentinus edodes (named Leras cDNA) can functionally replace its homolog genes (ScRAS1 and ScRAS2) in the yeast Saccharomyces cerevisiae to maintain the viability of an yeast strain containing genetic disruptions of both RAS genes. The strain replaced by a Leras-cDNA-carrying plasmid, however, grew slower than the strains replaced by a ScRAS1- or a ScRAS2-carrying plasmid. The intracellular level of cAMP in the strain harboring the Leras-cDNA-carrying plasmid was clearly higher than that of a parental strain which maintains a plasmid carrying the S. cerevisiae cAMP-dependent protein kinase catalytic subunit C1 gene, TPK1, but was lower than that in a strain harboring an ScRAS2-carrying plasmid. These results suggest that the Leras cDNA can complement the ras1- ras2- mutation of yeast by virture of the stimulation of adenylate cyclase activity, although the complementation is not as efficient as that obtained by expressing the ScRAS2 gene.
通过质粒交换程序表明,担子菌香菇的Ras编码cDNA(命名为Leras cDNA)可以在功能上替代酿酒酵母中的同源基因(ScRAS1和ScRAS2),以维持含有两个RAS基因遗传破坏的酵母菌株的活力。然而,被携带Leras-cDNA的质粒替代的菌株比被携带ScRAS1或ScRAS2的质粒替代的菌株生长得慢。携带Leras-cDNA的质粒的菌株中cAMP的细胞内水平明显高于维持携带酿酒酵母cAMP依赖性蛋白激酶催化亚基C1基因TPK1的质粒的亲本菌株,但低于携带ScRAS2的质粒的菌株。这些结果表明,Leras cDNA可以通过刺激腺苷酸环化酶活性来补充酵母的ras1- ras2-突变,尽管这种互补不如通过表达ScRAS2基因获得的互补有效。