Ahmad D, Massé R, Sylvestre M
Centre de Recherche en Microbiologie Appliquée, Institut Armand-Frappier, Université du Québec, Laval, Canada.
Gene. 1990 Jan 31;86(1):53-61. doi: 10.1016/0378-1119(90)90113-6.
The genes of Pseudomonas testosteroni strain B-356, specifying the transformation of 4-chlorobiphenyl (4-CB) into 4-chlorobenzoic acid (4-CBA) were cloned into Pseudomonas putida KT2440 using a broad-host-range cosmid, pPSA842. Of 10,000 clones tested, four were able to transform 4-CB. Gas chromatographic and mass spectrometric analysis of the catabolic products from two of the 4-CB-transforming clones carrying the hybrid plasmids, pDA1 and pDA2, demonstrated that pDA1 carried a complete set of structural genes involved in the transformation of 4-CB into 4-CBA, while pDA2 contained part of the pathway genes leading up to the meta-cleavage compound. Restriction endonuclease mapping and subcloning of pDA1 and pDA2 showed that the clones contained a common stretch of DNA of about 9.1 kb and that pDA2 carried gene(s) involved in regulation. Probing blots of genomic DNA from 13 different polychlorinated biphenyl(PCB)-degrading bacteria with radio-labelled pDA1 and pDA2, suggested that many PCB-degrading pathways have a common phylogenetic origin.
利用广宿主黏粒pPSA842，将睾丸酮假单胞菌B-356菌株中负责将4-氯联苯（4-CB）转化为4-氯苯甲酸（4-CBA）的基因克隆到恶臭假单胞菌KT2440中。在测试的10000个克隆中，有4个能够转化4-CB。对携带杂交质粒pDA1和pDA2的两个4-CB转化克隆的分解代谢产物进行气相色谱和质谱分析表明，pDA1携带了一组完整的将4-CB转化为4-CBA的结构基因，而pDA2包含了通向间位裂解化合物的部分途径基因。pDA1和pDA2的限制性内切酶图谱分析和亚克隆表明，这些克隆含有一段约9.1 kb的共同DNA片段，并且pDA2携带参与调控的基因。用放射性标记的pDA1和pDA2对13种不同的多氯联苯（PCB）降解细菌的基因组DNA印迹进行杂交，表明许多PCB降解途径具有共同的系统发育起源。
