Bacilysin overproduction in Bacillus amyloliquefaciens FZB42 markerless derivative strains FZBREP and FZBSPA enhances antibacterial activity
Bacilysin overproduction in Bacillus amyloliquefaciens FZB42 markerless derivative strains FZBREP and FZBSPA enhances antibacterial activity
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解淀粉芽孢杆菌 FZB42 无标记衍生菌株 FZBREP 和 FZBSPA 中杆菌肽过量产生增强抗菌活性
DOI：
10.1007/s00253-014-6251-0
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发表时间：
2014-12
影响因子：
5
通讯作者：
Gao Xuewen
中科院分区：
文献类型：
--
作者：
Wu Liming;Wu Huijun;Chen Lina;Lin Ling;Rainer Borriss;Gao Xuewen
解淀粉芽孢杆菌菌株FZBREP和FZBSPA通过分别用来自质粒pMK 3和pLOSS的组成型启动子P repB和P spac替换天然杆菌溶素操纵子启动子而衍生自野生型FZB 42。这些菌株含有两个抗生素抗性基因，并且通过使用从温度敏感载体pLOSS-cre表达的Cre重组酶删除氯霉素抗性盒和与两个lox位点（lox 71和lox 66）邻接的启动子区域来构建无标记菌株。通过高温（50 °C）处理去除载体编码的壮观霉素抗性基因。RT-PCR和qRT-PCR结果表明，PrepB，尤其是Pspac能显著提高bac操纵子的表达量，FZBREP和FZBSPA菌株产生的杆菌溶素分别比野生型高170.4%和315.6%。芽孢杆菌溶素的过量产生伴随着对金黄色葡萄球菌（芽孢杆菌溶素的指示物）和密歇根棒杆菌亚种的拮抗活性的增强。sepedonicum（马铃薯环腐病的病原体）。与野生型菌株相比，环腐病相关的坏死块茎的大小和程度都有所降低，这证实了这些基因工程菌株的保护作用和生防潜力。
Bacillus amyloliquefaciens strains FZBREP and FZBSPA were derived from the wild-type FZB42 by replacement of the native bacilysin operon promoter with constitutive promoters P repB and P spac from plasmids pMK3 and pLOSS, respectively. These strains contained two antibiotic resistance genes, and markerless strains were constructed by deleting the chloramphenicol resistance cassette and promoter region bordered by two lox sites (lox71 and lox66) using Cre recombinase expressed from the temperature-sensitive vector pLOSS-cre. The vector-encoded spectinomycin resistance gene was removed by high temperature (50 °C) treatment. RT-PCR and qRT-PCR results indicated that P repB and especially P spac significantly increased expression of the bac operon, and FZBREP and FZBSPA strains produced up to 170.4 and 315.6% more bacilysin than wild type, respectively. Bacilysin overproduction was accompanied by enhancement of the antagonistic activities against Staphylococcus aureus (an indicator of bacilysin) and Clavibacter michiganense subsp. sepedonicum (the causative agent of potato ring rot). Both the size and degree of ring rot-associated necrotic tubers were decreased compared with the wild-type strain, which confirmed the protective effects and biocontrol potential of these genetically engineered strains.
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影响因子：
2
作者：
R. Dmochowski
通讯作者：
R. Dmochowski
DOI：
10.1016/j.respe.2020.09.004
发表时间：
2020-10-10
期刊：
Revue D'Epidemiologie et De Sante Publique
影响因子：
--
作者：
La rédaction
通讯作者：
La rédaction
影响因子：
9.3
作者：
通讯作者：
--
影响因子：
5.9
作者：
Craighead FL
通讯作者：
Craighead FL