Weisshaar M P, Franklin F C, Reineke W
J Bacteriol. 1987 Jan;169(1):394-402. doi: 10.1128/jb.169.1.394-402.1987.
The genes specifying the utilization of 3-chlorobenzoate by Pseudomonas sp. strain B13 WR1 have been cloned by using a broad-host-range cosmid cloning system. Analysis of the catabolic products of the enzymatic reactions encoded by two hybrid cosmids, pMW65 and pMW90, by thin-layer and high-performance liquid chromatography demonstrated that both encoded the genes for the complete catabolism of 3-chlorobenzoate. Physical analysis of one of the cosmid derivatives, pMW65, by restriction endonuclease mapping and subcloning demonstrated that the pathway genes are encoded on a fragment no larger than 11 kilobases.
利用广宿主范围黏粒克隆系统,已克隆出假单胞菌属菌株B13 WR1中指定利用3 - 氯苯甲酸的基因。通过薄层色谱法和高效液相色谱法对两个杂种黏粒pMW65和pMW90所编码的酶促反应的分解代谢产物进行分析,结果表明二者均编码3 - 氯苯甲酸完全分解代谢的基因。通过限制性内切酶图谱分析和亚克隆对其中一个黏粒衍生物pMW65进行物理分析,结果表明该途径基因位于一个不大于11千碱基的片段上。