Establishment of electroporation protocol for Bacillus subtilis 1012 and WB800N

Abstract

Bacillus subtilis has been developed as an attractive expression host because of many advantages. For examples, it is nonpathogenic and allows secretion of functional extracellular proteins directly into the culture medium; about 60 % of industrial enzymes available produced by Bacillus species. To use B. subtilis strain for research and as host strain for expression of recombinant protein, bacterial genetic methods should be developed. Electroporation to transfer directly DNA into B. subtilis is one of the methods that draw a lot of attention of scientists. A problem encountered in the methods that draw a lot of attention of scientists. A problem encountered in the electroporation of DNA into B. subtilis is that an established protocol for one strain can hardly be used for another strain. B. subtilis 1012 and WB800N have recently been used as expression hosts for expression of recombinant proteins, but electroporation method has not been established. In this study, we use a pHT plasmid to establish an electroporation protocol for B. subtilis 1012 and WB800N. The influence of sampling time, concentration and time for incubating with lysozyme, voltage on the transformation was investigated to establish the protocol.