Tsoĭ T V, Kosheleva I A, Zamaraev V S, Trelina O V, Selifonov S A
Genetika. 1988 Sep;24(9):1550-61.
The genes nahC and nahD from Pseudomonas putida naphthalene degradation plasmid pBS286 were cloned on the vector pUC19 in Escherichia coli cells. The catechol-2,3-oxygenase activity observed in E. coli cells containing recombinant plasmid pBS955 demands the participation of 32 kD polypeptide which is apparently the product of the nahC gene. Second polypeptide of molecular weight 34.5 kD is synthesized in pBS955 containing E. coli minicells and perhaps it is a nahD gene product. The data obtained indicate that 1,2-dihydroxynaphthalene dioxygenase possesses a relaxed substrate specificity, at least, when cloned on a multicopy vector. The cloned DNA insert of pBS955 is not likely to contain its own promoter, so that expression of nahC and nahD genes from the nah1 operon is controlled by the vector lac promoter. The direction of transcription and localization of both polypeptides on the pBS955 map are determined.
恶臭假单胞菌萘降解质粒pBS286上的基因nahC和nahD在大肠杆菌细胞中克隆到载体pUC19上。在含有重组质粒pBS955的大肠杆菌细胞中观察到的儿茶酚-2,3-加氧酶活性需要32 kD多肽的参与，该多肽显然是nahC基因的产物。分子量为34.5 kD的第二条多肽在含有pBS955的大肠杆菌微小细胞中合成，它可能是nahD基因的产物。获得的数据表明，1,2-二羟基萘双加氧酶至少在克隆到多拷贝载体上时具有宽松的底物特异性。pBS955的克隆DNA插入片段不太可能含有其自身的启动子，因此来自nah1操纵子的nahC和nahD基因的表达由载体lac启动子控制。确定了两条多肽在pBS955图谱上的转录方向和定位。
