CONSTRUCTION OF LYS2 CARTRIDGES FOR USE IN GENETIC MANIPULATIONS OF SACCHAROMYCES-CEREVISIAE
CONSTRUCTION OF LYS2 CARTRIDGES FOR USE IN GENETIC MANIPULATIONS OF SACCHAROMYCES-CEREVISIAE
复制标题
DOI：
10.1016/0378-1119(86)90408-7
复制
发表时间：
1986-01-01
期刊：
影响因子：
3.5
通讯作者：
PHILIPPSEN, P
中科院分区：
文献类型：
--
作者：
FLEIG, UN;PRIDMORE, RD;PHILIPPSEN, P
将接头阵列添加到酿酒酵母LYS2基因的5“和3”边界，这允许产生具有不同粘性末端的18个LYS2盒。因为它是必要的，以确定开始和结束的近似。4.5kb的LYS2基因，我们测定了其5''区的1kb和3''区的1.5kb，并定位了mRNA起始点。通过将推测的ATG上游序列与S.酿酒酵母CYC1启动子和随后的体内LYS2表达的证明。通过使用配有LYS2盒的质粒将lys2突变株转化为Lys+原养型，证明了LYS2盒的正常功能。
Linker arrays were added to the 5'' and 3'' boundaries of the Saccharomyces cerevisiae LYS2 gene, which allow the generation of 18 LYS2 cartridges with different sticky ends. As it was necesary to define the beginning and the end of the approx. 4.5-kb LYS2 gene, we sequenced 1 kb of its 5'' and 1.5 kb of its 3'' region and mapped the mRNA start point. The open reading frame (ORF) found by this analysis was proven to the LYS2 ORF by exchanging the sequences upstream from the presumptive ATG with the S. cerevisiae CYC1 promoter and subsequent demonstration of LYS2 expression in vivo. The proper functioning of the LYS2 cartridges was demonstrated by the transformation of lys2 mutant strains to Lys+ prototrophy using plamids furnished with a LYS2 cartridge.