title: Artificial induction of genetic competence in Bacillus amyloliquefaciens isolates.
abstract: OBJECTIVE
  To induce natural genetic competence in Bacillus amyloliquefaciens isolates through overexpression of the master regulator, ComK, from B. subtilis (ComKBsu).
  
  
  RESULTS
  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 ± 20.6 to 1.7 ± 0.1 × 105cfu (colony-forming units) per μg DNA, and the transformation efficiencies of PCR-assembled deletion constructs ranged from 3.2 ± 0.76 to 3.5 ± 0.42 × 104cfu per μg DNA in the four tested strains.
  
  
  CONCLUSIONS
  Artificial induction of genetic competence through overexpressing ComKBsuin B. amyloliquefaciens completed the tasks of replicative plasmid delivery and gene knockout via direct transformation of PCR-generated deletion cassettes.
authors: Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.
authors: Xue-Ting
authors: XT
authors: Chen
authors: Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.
authors: Jun-Bin
authors: JB
authors: Ji
authors: Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.
authors: Yong-Chuang
authors: YC
authors: Liu
authors: Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.
authors: Bin
authors: B
authors: Ye
authors: Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.
authors: Chao-Yang
authors: CY
authors: Zhou
authors: Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China. yanxin@njau.edu.cn.
authors: Xin
authors: X
authors: Yan
mesh_terms: D000069977:Bacillus amyloliquefaciens / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D010957:Plasmids / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins
keywords: Bacillus amyloliquefaciens; ComK; Genetic competence; Genetic manipulation; Replicative plasmid delivery; Xylose