Griffin H G, Gasson M J
Genetics and Microbiology Department, AFRC Institute of Food Research, Colney, Norwich, UK.
Lett Appl Microbiol. 1993 Aug;17(2):92-6. doi: 10.1111/j.1472-765x.1993.tb00379.x.
Translational gene fusions between the Escherichia coli beta-galactosidase (lacZ) gene and the Lactococcus lactis lactose operon were constructed such that transcription from the lactose operon promoter could be assessed by measuring beta-galactosidase activity. The level of beta-galactosidase activity was up to 2.5-fold lower when MG5267 cells, which contain a chromosomal copy of the lactose operon, were grown in glucose compared to those grown in lactose. A greater degree of repression was seen in cells containing the multi-copy plasmid-encoded repressor than in those with only the single-copy chromosomal gene, indicating that the repressor protein is at least partly responsible for the reduction in expression when the cells are grown in glucose (i.e. in the absence of inducer). However, the beta-galactosidase activity was found to be 5.5-fold lower in glucose than in lactose in cells which lacked a fully functional lactose operon. The decrease in expression was shown to be due to glucose repression. The levels of expression when the cells were grown in glucose were considerably higher for MG5267 than for MG1363 suggesting perhaps that a product of the chromosomally-encoded operon in MG5267 has a positive effect on transcription.
构建了大肠杆菌β-半乳糖苷酶(lacZ)基因与乳酸乳球菌乳糖操纵子之间的翻译基因融合体,以便通过测量β-半乳糖苷酶活性来评估乳糖操纵子启动子的转录情况。与在乳糖中生长的MG5267细胞相比,含有乳糖操纵子染色体拷贝的MG5267细胞在葡萄糖中生长时,β-半乳糖苷酶活性水平低至2.5倍。与仅含有单拷贝染色体基因的细胞相比,含有多拷贝质粒编码阻遏物的细胞中观察到更高程度的阻遏,这表明当细胞在葡萄糖中生长(即不存在诱导物)时,阻遏蛋白至少部分导致了表达的降低。然而,在缺乏完全功能性乳糖操纵子的细胞中,发现葡萄糖中的β-半乳糖苷酶活性比乳糖中的低5.5倍。结果表明表达的降低是由于葡萄糖阻遏。MG5267细胞在葡萄糖中生长时的表达水平比MG1363高得多,这可能表明MG5267中染色体编码操纵子的产物对转录有积极影响。