I want to get a super bacteria with multiple characters like; phosphate solubilization, ACC deaminase activity, IAA production. The bacterial strain in which I want to incorporate these genes is Bacillus subtilis strain with antifungal activity. Is it possible to do so?
Of course you can. Take care of codon optimization (e.g. use synthetic genes if necessary), selecting good ( inducible) promoters; make a choice between expression from plasmids or via integration of constructs into the genome. You can consider putting genes in artificial operons that you can first construct in E.coli and then bring to Bacillus. Then you can coexpress them from one promoter. It helps if you can use natural competence of this Bacillus subtilis strain. Hope this helps,
Oscar Kuipers
I think theoretically it is possible ... you could try and tell us your success story...
Incorporating multiple characters won't be easy though doable theoretically. you need to have multiple compatible plasmids with different promoters. follow the link http://parts.igem.org/Bacillus_subtilis. here you will find multiple plasmids and promoters available for Bacillus. you can also choose to insert the genes into the chromosome. then also you need to be careful with the promoters used.
This way was used at 1990s (http://bashanis.org/gmaweb/pdfs/geneticmanip.pdf). We have tried to do it at 2000s with ISTC support, but there are many legal problem on this way at least in Russia and EU.
Dear Oscar Kuipers, your answer is very encouraging & helpful, thanks a lot. Thanks to all others for helpful answers. Since it was only idea to me, now going to structure it by Allah's Will. I would seek further guidance from you in future.
Alex if you can share information how you worked on it in detail. What about your experiment, was it successful? If you would like to share, you can share here or email me at [email protected]
Sound good, but it is tricky as these traits are result of complex process. Consider following important factors
1. Codon, promoter, reading frame, operon etc.
2. Plasmid load if you are express these genes through plasmid.
3. Selection of recombinant
4. Transportation of product outside the cell, taken care by signal peptide
Yes you can do it. First of all synthesize DNA strands having all these characters. However, study the strain on which you wish to carry out the engineering, to see how many of the characters (you are interested is already there on the genome). As I see your interest, you only need to make the organism synthesize IAA. That too is not a very difficult job. If you carefully make the protocol for your research methodology, you will find your work very easy.
Dear Jai
I have done through screening of my bacterial strain. This strain is unable to synthesize IAA, neither it solubilize insoluble phosphate. rather it has strong ability to control fungus. I need to insert atleast two characters i.e. IAA production & P-solubilization.
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