Tao Mei-Feng, Zhou Xiu-Fen, Kieser Tobias, Deng Zi-Xin
Key Laboratory of Agricultural Microbiology, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China.
Sheng Wu Gong Cheng Xue Bao. 2002 Jul;18(4):420-3.
pHZ1080, an E. coli-Streptomyces shuttle expression vector was constructed in order to explore the utilization of lambda phage regulated expression elements in Streptomyces. A 2.7 kb polyketide synthase (PKS) gene from Streptomyces sp. FR-008 was inserted into downstream of lambda phage promoter (PR) to give the shuttle plasmid, pHZ1067. The PKS protein was expressed in Streptomyces lividans carrying pHZ1067 in a heat-dependent manner, as it did in E. coli. The PKS protein expressed in both hosts with same molecular weight was detected by SDS-PAGE and Western-blot. The successful heat-induced expression of PKS suggested that pHZ1080 was useful and convenient for heat-induced expression of heterologous genes in both E. coli and Streptomyces.
构建了大肠杆菌-链霉菌穿梭表达载体pHZ1080，以探索λ噬菌体调控表达元件在链霉菌中的应用。将来自链霉菌FR-008的2.7 kb聚酮合酶（PKS）基因插入λ噬菌体启动子（PR）下游，得到穿梭质粒pHZ1067。PKS蛋白在携带pHZ1067的变铅青链霉菌中以热依赖的方式表达，如同在大肠杆菌中一样。通过SDS-PAGE和Western印迹检测到在两种宿主中表达的PKS蛋白分子量相同。PKS的成功热诱导表达表明pHZ1080对于在大肠杆菌和链霉菌中热诱导表达异源基因是有用且方便的。
