Brabetz W, Liebl W, Schleifer K H
Lehrstuhl für Mikrobiologie, Technische Universität München, Federal Republic of Germany.
Arch Microbiol. 1991;155(6):607-12. doi: 10.1007/BF00245357.
The entire Escherichia coli lactose operon was inserted into an E. coli/Corynebacterium glutamicum shuttle vector and introduced into the gram-positive host organism C. glutamicum R 163. Recombinant C. glutamicum strains carrying the lac genes downstream of an efficient promoter displayed rapid growth with lactose as the sole source of carbon. Two prerequisites were necessary to obtain good growth of C. glutamicum R 163 on lactose: presence of the lacY gene in addition to lacZ and an appropriate promoter for efficient transcription in C. glutamicum. The galactose moiety of lactose was not utilized but accumulated in the culture broth. C. glutamicum strains carrying only the lacZ (beta-galactosidase) gene but not lacY (lactose permease) were not able to grow in lactose minimal medium.
将完整的大肠杆菌乳糖操纵子插入到大肠杆菌/谷氨酸棒杆菌穿梭载体中,并导入革兰氏阳性宿主菌谷氨酸棒杆菌R 163。携带位于高效启动子下游的lac基因的重组谷氨酸棒杆菌菌株,以乳糖作为唯一碳源时生长迅速。要使谷氨酸棒杆菌R 163在乳糖上良好生长,有两个先决条件:除了lacZ基因外还需存在lacY基因,以及在谷氨酸棒杆菌中进行有效转录的合适启动子。乳糖的半乳糖部分未被利用而是积累在培养液中。仅携带lacZ(β-半乳糖苷酶)基因而不携带lacY(乳糖通透酶)基因的谷氨酸棒杆菌菌株,无法在乳糖基本培养基中生长。