Construction of a plasmid interspecific transfer system in Bacillus species with the counterselectable marker mazF

Joint Event on International Conference on Cancer Research & Diagnostics & 16^th Asia Pacific Biotechnology Congress

August 15-16, 2018 Singapore

Hui Song

TLI Foundation, China

Posters & Accepted Abstracts: J Tissue Sci Eng

Abstract :

Bacillus species strains as attractive hosts to produce heterologous secretory proteins usually play important roles in bioindustry. However, low transformation efficiency of exogenous plasmids limited the application of Bacillus species. Here, a novel plasmid interspecific transfer system, with higher transformation efficiency, higher positive rate and more convenient manipulation has been successfully constructed. A high transformation efficiency strain Bacillus subtilis F-168 containing the counter-selectable marker mazF was used as the plasmid donor strain in this transfer method. A shuttled plasmid pBE980b and its recombinant plasmids pBE980b::BapuI and pBE980::Spro1 were successfully transferred into the recipient Bacillus strains (Bacillus amyloliquefaciens 66, Bacillus licheniformis 124 and Bacillus megaterium 258) by this method. After co-culturing the donor cells (OD 600 nm=1.4-1.7) and the recipient cells (OD 600 nm=0.5-0.9) cells for 24 hours in 22oC, more than 1.0Ã?105 positive transformants were obtained and a transformation efficiency of 1.0Ã?10-3 showed 10-100 times higher than those transferred using traditional transfer methods. This plasmid interspecific transfer system would provide a new approach for genetic manipulation in Bacillus strains and accelerate the research progress of Bacillus strains in bio-industry.

Biography :

E-mail: song_h@tib.cas.cn