Large plasmid pCAR2 and class II transposon Tn4676 are functional mobile genetic elements to distribute the carbazole/dioxin-degradative car gene cluster in different bacteria
Large plasmid pCAR2 and class II transposon Tn4676 are functional mobile genetic elements to distribute the carbazole/dioxin-degradative car gene cluster in different bacteria
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DOI：
10.1007/s00253-004-1778-0
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发表时间：
2005-05-01
影响因子：
5
通讯作者：
Nojiri, H
中科院分区：
文献类型：
--
作者：
Shintani, M;Yoshida, T;Nojiri, H
对从食树脂假单胞菌 CA10 菌株中分离的咔唑分解代谢质粒 pCAR1 进行了完整测序；发现pCAR1携带含有咔唑降解基因的II类转座子Tn4676。在这项研究中，从恶臭假单胞菌菌株 HS01 中分离出一种名为 pCAR2 的新质粒，该菌株是咔唑降解菌假单胞菌之间交配的转接合子。菌株K23和恶臭假单胞菌菌株DS1。 pCAR1 和 pCAR2 的 Southern 杂交和核苷酸序列分析表明，两者的整个主链结构非常相似。质粒pCAR2是可自我传播的，因为它以每个受体细胞2x10(-7)的频率从菌株HS01转移到荧光假单胞菌菌株IAM12022。在丰富培养基上连续转移菌株HS01后，我们检测到Tn4676从pCAR2转座到HS01染色体。 Tn4676 的染色体定位副本的两侧是 6 bp 靶标重复 5'-AACATC-3'。这些结果通过实验证明了 pCAR2 的可转移性以及 Tn4676 在 pCAR2 上的功能。清楚地表明，质粒pCAR2和转座子Tn4676是活性可移动遗传元件，可以介导咔唑分解代谢基因的水平转移。
The carbazole-catabolic plasmid pCAR1 isolated from Pseudomonas resinovorans strain CA10 was sequenced in its entirety; and it was found that pCAR1 carries the class II transposon Tn4676 containing carbazole-degradative genes. In this study, a new plasmid designated pCAR2 was isolated from P. putida strain HS01 that was a transconjugant from mating between the carbazole-degrader Pseudomonas sp. strain K23 and P. putida strain DS1. Southern hybridization and nucleotide sequence analysis of pCAR1 and pCAR2 revealed that the whole backbone structure was very similar in each. Plasmid pCAR2 was self-transmissible, because it was transferred from strain HS01 to P. fluorescens strain IAM12022 at the frequency of 2x10(-7) per recipient cell. After the serial transfer of strain HS01 on rich medium, we detected the transposition of Tn4676 from pCAR2 to the HS01 chromosome. The chromosome-located copy of Tn4676 was flanked by a 6-bp target duplication, 5'-AACATC-3'. These results experimentally demonstrated the transferability of pCAR2 and the functionality of Tn4676 on pCAR2. It was clearly shown that plasmid pCAR2 and transposon Tn4676 are active mobile genetic elements that can mediate the horizontal transfer of genes for the catabolism of carbazole.