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外膜囊泡介导大肠杆菌中好氧反硝化基因的水平转移。

Outer membrane vesicles mediated horizontal transfer of an aerobic denitrification gene between Escherichia coli.

机构信息

Department of Environmental Engineering, College of Biology and the Environment, Nanjing Forestry University, Nanjing, 210037, China.

Nanjing Yi Wei Environmental Protection Technology Co., Ltd., Nanjing, 210048, China.

出版信息

Biodegradation. 2021 Aug;32(4):435-448. doi: 10.1007/s10532-021-09945-y. Epub 2021 Apr 22.

Abstract

Bacterial genetic material can be horizontally transferred between microorganisms via outer membrane vesicles (OMVs) released by bacteria. Up to now, the application of vesicle-mediated horizontal transfer of "degrading genes" in environmental remediation has not been reported. In this study, the nirS gene from an aerobic denitrification bacterium, Pseudomonas stutzeri, was enclosed in a pET28a plasmid, transformed into Escherichia coli (E. coli) DH5α and expressed in E. coli BL21. The E. coli DH5α released OMVs containing the recombination plasmid pET28a-nirS-EGFP. When compared with the free pET28a-nirS-EGFP plasmid's inability to transform, nirS in OMVs could be transferred into E. coli BL21 with the transformation frequency of 2.76 × 10 CFU/g when the dosage of OMVs was 200 µg under natural conditions, and nirS could express successfully in recipient bacteria. Furthermore, the recipient bacteria that received OMVs containing pET28a-nirS-EGFP could produce 18.16 U/mL activity of nitrite reductase.

摘要

细菌的遗传物质可以通过细菌释放的外膜囊泡(OMVs)在微生物之间进行水平转移。到目前为止,还没有报道过利用囊泡介导的“降解基因”在环境修复中的水平转移。在本研究中,来自好氧反硝化细菌恶臭假单胞菌的nirS 基因被包裹在 pET28a 质粒中,转化到大肠杆菌(E. coli)DH5α 中,并在大肠杆菌 BL21 中表达。E. coli DH5α 释放出含有重组质粒 pET28a-nirS-EGFP 的 OMVs。与游离的 pET28a-nirS-EGFP 质粒相比,在自然条件下,当 OMVs 的剂量为 200µg 时,OMVs 中的 nirS 可以以 2.76×10 CFU/g 的转化频率转移到大肠杆菌 BL21 中,并且 nirS 可以在受体细菌中成功表达。此外,接收含有 pET28a-nirS-EGFP 的 OMVs 的受体细菌可以产生 18.16 U/mL 的亚硝酸盐还原酶活性。

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