Sarthy A V, McConaughy B L, Lobo Z, Sundstrom J A, Furlong C E, Hall B D
Department of Genetics, University of Washington, Seattle 98195.
Appl Environ Microbiol. 1987 Sep;53(9):1996-2000. doi: 10.1128/aem.53.9.1996-2000.1987.
Transformation of Saccharomyces cerevisiae by yeast expression plasmids bearing the Escherichia coli xylose isomerase gene leads to production of the protein. Western blotting (immunoblotting) experiments show that immunoreactive protein chains which comigrate with the E. coli enzyme are made in the transformant strains and that the amount produced parallels the copy number of the plasmid. When comparable amounts of immunologically cross-reactive xylose isomerase protein made in E. coli or S. cerevisiae were assayed for enzymatic activity, however, the yeast protein was at least 10(3)-fold less active.
携带大肠杆菌木糖异构酶基因的酵母表达质粒对酿酒酵母进行转化后会产生该蛋白质。蛋白质印迹法(免疫印迹法)实验表明,在转化菌株中产生了与大肠杆菌酶迁移率相同的免疫反应性蛋白链,并且产生的量与质粒的拷贝数平行。然而,当对在大肠杆菌或酿酒酵母中产生的等量具有免疫交叉反应性的木糖异构酶蛋白进行酶活性测定时,酵母蛋白的活性至少低10³倍。