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