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Marking Bacillus sublitis with GFP


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#1 smallcat227

smallcat227

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Posted 02 July 2009 - 10:07 PM

Hello everyone,

We want to use green fluorescence protein as molecular marker for tagging Bacillus subtilis in soil. We used plasmid pUBC19 to construct the shuttle vector, the promotor was screened in the genome of the Bacillus strains. The GFP shuttle vector can express well in the E.coli, but when we transform them into the Bacillus, whether electrotransformation or chemicaltransformation, the express level of the GFP was very low, under the microscope only very weak fluorescence can be seen.

We have trid many promotors but can not help, also we can make sure that the shuttle vector has been transformed into the Bacillus, everyone who can tell us what's the matter, the promotor was not strong enough, or the GFP has been hydrolyze by the protease, or other reasons? And what should we do next?

Thanks a lot!




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