Olekhnovich I N, Fomichev Y K
Department of Microbiology, Belarus State University, Minsk.
Gene. 1994 Mar 11;140(1):63-5. doi: 10.1016/0378-1119(94)90731-5.
A cloning vector pPS7 (8.5 kb) for Pseudomonas was constructed from pBR322 and the Pseudomonas cryptic low-copy-number pMK1 plasmid. The vector confers resistance to kanamycin (Km) and tetracycline (Tc), contains the par locus of Pseudomonas plasmid pMT2 and a mob site. The new vector was used for construction of controlled-expression vector pPS10 (10.4 kb) based on the trpIBA genes of Pseudomonas putida. This KmR vector contains the trpI gene, encoding activator protein and promoter of trpBA genes (Pba), which are inducible by TrpI and indoleglycerol phosphate (InGP). InGP is an unstable compound, but it accumulates in trpE mutants grown in anthranilate (Anth)-supplemented medium. We show that expression of the Escherichia coli pheA gene, inserted into the pPS10 vector downstream from Pba, increases about 70-fold upon InGP accumulation.
一种用于假单胞菌的克隆载体pPS7（8.5 kb）由pBR322和假单胞菌隐蔽型低拷贝数pMK1质粒构建而成。该载体赋予对卡那霉素（Km）和四环素（Tc）的抗性，含有假单胞菌质粒pMT2的par位点和一个mob位点。新载体用于基于恶臭假单胞菌的trpIBA基因构建可控表达载体pPS10（10.4 kb）。这个KmR载体包含trpI基因，其编码激活蛋白以及trpBA基因（Pba）的启动子，它们可被TrpI和吲哚甘油磷酸（InGP）诱导。InGP是一种不稳定的化合物，但它在添加邻氨基苯甲酸（Anth）的培养基中生长的trpE突变体中积累。我们表明，插入到pPS10载体中Pba下游的大肠杆菌pheA基因的表达，在InGP积累时增加约70倍。
