[DIYbio] Bacillus: An alternative for E coli in DIYbio?

Hey guys!

Recently I have begun using Bacillus subtilis a lot, and I gotta say... it is a LOT easier then E coli. For example I did one experiment with a strain of Bacillus that over expressed comK under a xylose inducible promoter... all I did was get the cells in growth phase, give them some DNA, grow them for 45 minutes then plate. I got transformation efficiency of 6x 10^3 with a circular integration plasmid. That is even easier then Cathal's Bacillus transformation protocol (no offense (:  ), which is one of the simplest out there. I can easily get better expression of comK by optomizing the RBS and I can make this work under  a sucrose inducible promoter! (Working on that right now) In addition to this, I have found a way to negatively select in Bacillus (I think it is the mazF gene from e coli), meaning that I could possibly make a strain which all you need to do is transform them, no selection method needed! 

Plus Bacillus can take pure PCR fragments... meaning that using homologous recombination and negative selection you could literally PCR an interesting protein from lets say, some cheek cells, then integrate it into bacillus. The integration could then be expressed, secreted, and then purified (using a variety of tags already in the bacillus, given that the gene has no introns/exons) Skip plasmids and e coli entirely! (of course... I need to make these strains)

Anyone have anything to add or conflicting views?

-Koeng

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