Electroporation is a feasible method to introduce circularized or linearized DNA into B. subtilis chromosome.
Here, we present the electroporation as a feasible and efficient method for introducing circularized and linearized DNA into Bacillus subtilis chromosome. Two integration experiments were carried out and demonstrated the feasibility and efficiency of electroporation to introduce the target DNA into the B. subtilis chromosome. By using of electroporation, a multiple-cistron contained five genes from B. subtilis biotin biosynthetic pathway was introduced into the B. subtilis chromosome efficiently and created a repeated copy in chromosome via a single crossover event. Then an ectopic promoter was introduced conveniently into the upstream of one of the repeated multiple-cistron via a double crossover event. To further demonstrate the application of electroporation in genetic research, the early sporulation gene spo0A of B. subtilis was knocked out and, consequently, the null of sporulation and logged growth was observed in this study. Thus, the electroporation as an alternative method of integration in B. subtilis is feasible and practical.
Yang
Ming-Ming
MM
College of Animal Sciences, Northwest A&F University, 712100, Yangling, People's Republic of China. yangjinxin@vip.163.com
Zhang
Wei-Wei
WW
Bai
Xiao-Ting
XT
Li
Heng-Xin
HX
Cen
Pei-Lin
PL
D001412:Bacillus subtilis / Q000235:genetics* / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D004270:DNA, Circular / Q000235:genetics*; D018274:Electroporation / Q000379:methods*; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D005798:Genes, Bacterial / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D015203:Reproducibility of Results; D013171:Spores, Bacterial / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; C080950:Spo0A protein, Bacillus subtilis; D014157:Transcription Factors