Abstract
Objective To explore an effective technique to control the horizontal transfer of drug-resistance genes using specific CRISPR/Cas9 system. Methods Three pairs of kanamycin-resistance gene (Kan)-specific single guide RNAs (sgRNAs) were designed, annealed and inserted into the Bsa Ⅰ site of pCas9, respectively. Each group of recombinant bacteria competent cells were transformed by pET-30a that carried Kan gene. The transformants were spread on plates containing chloramphenicol or chloramphenicol/kanamycin to calculate and compare the transformation efficiencies. The DH5α competent cells containing pET-30a were transformed with each recombinant plasmid to detect the degradation of Kan gene mediated by specific sgRNAs with PCR and to observe the effects on growth curve of recombinant. Results The transformation efficiencies of Kan gene into Escherichia coli strains harboring specific sgRNAs CRISPR/Cas9 system decreased 100%-88.08%. The specific sgRNAs CRISPR/Cas9 system degraded Kan gene without affecting the growth of bacteria. Conclusion The specific CRISPR/Cas9 system inhibits the horizontal transfer of bacterial kanamycin-resistance gene. Key words: CRISPR/Cas9 system; Drug resistance; Horizontal gene transfer
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
