Artificial induction of genetic competence in Bacillus amyloliquefaciens isolates
Artificial induction of genetic competence in Bacillus amyloliquefaciens isolates
复制标题
人工诱导解淀粉芽孢杆菌分离株的遗传能力。
DOI：
10.1007/s10529-016-2194-0
复制
发表时间：
2016-12-01
影响因子：
2.7
通讯作者：
Yan, Xin
中科院分区：
文献类型：
--
作者：
Chen, Xue-Ting;Ji, Jun-Bin;Yan, Xin
为了通过过量表达枯草芽孢杆菌(B.subtilis，ComK(BSU))的主调控子comK来诱导淀粉芽孢杆菌的自然遗传能力，以pUB110为骨架构建了携带木糖诱导的comK表达框的质粒pUBXC。通过pLS20介导的双亲接合，可以将pUBXC从枯草芽孢杆菌转移到淀粉液化芽孢杆菌。经木糖诱导后，4株含有pUBXC质粒的淀粉液化芽孢杆菌产生了遗传活性。在优化条件下，4株供试菌株的DNA转化效率分别为129+/-A 20.6~1.7+/-A 0.1×10(5)cfu(菌落形成单位)，4个供试菌株的转化效率分别为3.2+/-A 0.76~3.5+/-A 0.42×10(4)cfu/mgDNA。
To induce natural genetic competence in Bacillus amyloliquefaciens isolates through overexpression of the master regulator, ComK, from B. subtilis (ComK (Bsu) ).Plasmid pUBXC carrying the xylose-inducible comK expression cassette was constructed using plasmid pUB110 as a backbone. Plasmid pUBXC could be transferred from B. subtilis to B. amyloliquefaciens through plasmid pLS20-mediated biparental conjugation. After being induced by xylose, four B. amyloliquefaciens strains harbouring plasmid pUBXC developed genetic competence. Under optimal conditions, the transformation efficiencies of plasmid DNA ranged from 129 +/- A 20.6 to 1.7 +/- A 0.1 x 10(5) cfu (colony-forming units) per mu g DNA, and the transformation efficiencies of PCR-assembled deletion constructs ranged from 3.2 +/- A 0.76 to 3.5 +/- A 0.42 x 10(4) cfu per mu g DNA in the four tested strains.Artificial induction of genetic competence through overexpressing ComK (Bsu) in B. amyloliquefaciens completed the tasks of replicative plasmid delivery and gene knockout via direct transformation of PCR-generated deletion cassettes.