Group II intron-anchored gene deletion in Clostridium

Kaizhi Jia, Yan Zhu, Yanping Zhang, Yin Li

Research output: Contribution to journalArticleResearchpeer-review

21 Citations (Scopus)

Abstract

Clostridium plays an important role in commercial and medical use, for which targeted gene deletion is difficult. We proposed an intron-anchored gene deletion approach for Clostridium, which combines the advantage of the group II intron "ClosTron" system and homologous recombination. In this approach, an intron carrying a fragment homologous to upstream or downstream of the target site was first inserted into the genome by retrotransposition, followed by homologous recombination, resulting in gene deletion. A functional unknown operon CAC1493-1494 located in the chromosome, and an operon ctfAB located in the megaplasmid of C. acetobutylicum DSM1731 were successfully deleted by using this approach, without leaving antibiotic marker in the genome. We therefore propose this approach can be used for targeted gene deletion in Clostridium. This approach might also be applicable for gene deletion in other bacterial species if group II intron retrotransposition system is established. 

Original languageEnglish
Article numbere16693
Number of pages10
JournalPLoS ONE
Volume6
Issue number1
DOIs
Publication statusPublished - 31 Jan 2011
Externally publishedYes

Keywords

  • introns
  • clostridium
  • gene targeting
  • deletion mutation
  • polymerase chain reaction
  • operons
  • homologous recombination
  • erythromycin

Cite this