Chevallier M R, Bloch J C, Lacroute F
Gene. 1980 Oct;11(1-2):11-9. doi: 10.1016/0378-1119(80)90082-7.
Chimeric plasmids composed of the bacterial plasmid pBR322, 2 micron yeast plasmid fragments and the 1.1 kb ura3+ fragment of yeast chromosomal DNA which codes for orotidine-5'-phosphate (OMP) decarboxylase were constructed and used to transform Escherichia coli and Saccharomyces cerevisiae recipient cells. The expression in yeast of one such plasmid was studied and compared to the expression of a chromosomally integrated bacterial plasmid. In the strain carrying the chimeric plasmid the level of OMP decarboxylase activity is about 25 times that found in either the wild-type strain or in the strain carrying the chromosomally integrated plasmid. The ampicillin gene of pBR322 is expressed in yeast. Labeling kinetics of RNA and measurements of the polyadenylated fractions showed that RNA hybridizing to the pBR322 plasmid was polyadenylated to the same extent as RNA hybridizing to the ura3+ gene. Half-lives of 10 and 20 min were estimated for the ura3+ and pBR322 transcripts respectively.
构建了由细菌质粒pBR322、2μm酵母质粒片段和酵母染色体DNA的1.1 kb ura3 +片段组成的嵌合质粒，该片段编码乳清苷-5'-磷酸（OMP）脱羧酶，并用于转化大肠杆菌和酿酒酵母受体细胞。研究了其中一种质粒在酵母中的表达，并与染色体整合细菌质粒的表达进行了比较。在携带嵌合质粒的菌株中，OMP脱羧酶活性水平约为野生型菌株或携带染色体整合质粒的菌株中发现的活性水平的25倍。pBR322的氨苄青霉素基因在酵母中表达。RNA的标记动力学和聚腺苷酸化部分的测量表明，与pBR322质粒杂交的RNA与与ura3 +基因杂交的RNA聚腺苷酸化程度相同。ura3 +和pBR322转录本的半衰期分别估计为10分钟和20分钟。
