Yang R, Hu Z, Deng Z, Li J
Huazhong Agricultural University, Wuhan, China.
Chin J Biotechnol. 1998;14(1):1-8.
pIJ4123 and pIJ6021 are high-copy-number vectors for gene expression in Streptomyces. They contain a strong, thiostrepton-inducible promoter, PtipA. Two E. coli-Streptomyces shuttle vectors, pHZ1271 and pHZ1272, were constructed by inserting a replicon and bla gene from E. coli into pIJ4123 and pIJ6021, respectively. Both vectors were structurally stable either in E. coli or in Streptomyces lividans. To demonstrate the utility of pHZ1272, the hemoglobin gene of Vitreoscillia (vhb) was cloned into pHZ1272 and expressed in Streptomyces lividans. The expression product (VHb) was detected by Western blotting and carbon monoxide binding activity analysis.
pIJ4123和pIJ6021是用于链霉菌基因表达的高拷贝数载体。它们含有一个强的、硫链丝菌素诱导型启动子PtipA。通过分别将来自大肠杆菌的复制子和bla基因插入pIJ4123和pIJ6021中，构建了两个大肠杆菌-链霉菌穿梭载体pHZ1271和pHZ1272。这两个载体在大肠杆菌或天蓝色链霉菌中结构都很稳定。为了证明pHZ1272的实用性，将透明颤菌血红蛋白基因（vhb）克隆到pHZ1272中并在天蓝色链霉菌中表达。通过蛋白质免疫印迹和一氧化碳结合活性分析检测到了表达产物（VHb）。
