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Outer membrane vesicles mediated horizontal transfer of an aerobic denitrification gene between Escherichia coli
Biodegradation ( IF 3.1 ) Pub Date : 2021-04-22 , DOI: 10.1007/s10532-021-09945-y
Weichuan Qiao 1, 2 , Lianjie Wang 1 , Yang Luo 1 , Jiahui Miao 1
Affiliation  

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 × 106 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.



中文翻译:

外膜囊泡介导有氧反硝化基因在大肠杆菌之间的水平转移

细菌遗传物质可以通过细菌释放的外膜囊泡(OMV)在微生物之间水平转移。迄今为止,还没有报道过囊泡介导的“降解基因”水平转移在环境修复中的应用。在这项研究中,NIRS从好氧反硝化细菌基因,施氏假单胞菌,被封闭在pET28a中的质粒,转化到大肠杆菌大肠杆菌)DH5α和在表达大肠杆菌BL21。该大肠杆菌含有重组DH5α释放OMV的质粒的pET28a NIRS -EGFP。与免费的pET28a– nirS相比-EGFP的质粒的无能变换,NIRS在OMV的可转移到大肠杆菌BL21与2.76转化频率×10 6  CFU /克时的OMV的用量为在自然条件下200微克,和近红外光谱可以在受体细菌成功表达。此外,接受含有pET28a– nirS -EGFP的OMV的受体细菌可以产生18.16 U / mL的亚硝酸还原酶活性。

更新日期:2021-04-22
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