ARS | Publication request: Electrotransformation of Bacillus mojavensis with fluorescent protein markers

Submitted to: Journal of Microbiological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 1, 2008
Publication Date: July 31, 2008
Repository URL: http://hdl.handle.net/10113/32758
Citation: Olubajo, B.A., Bacon, C.W. 2008. Electrotransformation of Bacillus mojavensis with fluorescent protein markers. Journal of Microbiological Methods. 74:102-105.

Interpretive Summary: We are interested in developing a biocontrol agent that will protect corn from pathogenic and toxic fungi. We have a species of bacteria that has the potential for controlling corn diseases but require a procedure to measure the interaction of this bacterium on corn plants during the period of protection. The approach required the marking of the bacterium with fluorescent genes that can be used to following the interaction of the bacterium in corn. This process referred to as transformation is a difficult procedure as bacteria of this group have not been transformed before. However, we report in this work the first successful transformation of this biocontrol species of bacterium, which was patented by the agency to protect corn and now will allow us to follow this bacterium and measured its interaction during the growth of corn throughout the season.

Technical Abstract: Gram-positive endophytic bacteria are difficult to transform. To study endophytic interactions between Bacillus mojavensis and maize, a method was developed to transform this species by electroporation with three fluorescent protein expressing integrative plasmids: pSG1154, pSG1192, and pSG1193. The transformations achieved were relatively high and verified by antibiotic selection, microscopy and amylase deficiency analyses. This represents the first successful transformation of this endophytic species.