An investigation of bacterial transformation techniques
Moldovan, Kimberly M.
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This thesis contain information regarding two possible, means of causing B. megaterium to take into , its, own genome # exogenous DNA. The whole goal of the transformation process, be it via electroporation or protoplast transformation, is to get the cell to not; only accept the foreign DNA, but to express the proteins the exogenous genes code for. The electroporation process consists of subjecting the bacterial cells to various strengths, of electric shock in order to cause pore formation in the celL-wall and subsequent entrance of the exogenous _DNA of interest. The disruption of the cell wall must not be too great that it is unable to synthesize new cell wall components and express the new genetic material. Protoplast transformation involves the complete removal of the bacterial cell wall, membrane fusion, membrane separation, cell-wall regeneration, and finally, expression of the new genetic matter. The former process would to be more efficient and successful, but per my research of one year and a half, it seems the exact opposite it true. This thesis investigated the parameters of both transformation processes and suggests future studies in this relatively new and exciting field of Microbial genetics. Included in the work are the parameters explored and several suggestions for further research. One of the more radical suggestions made about future studies is the concept of a fusion of techniques from both processes to increase transformation efficiencies. The concept is one of creating the cell wall-less protoplasts and add exogenous DNA, but then subject this to the electric shock.