Thesis (M.Sc.) - Cairo University - National Institute of Laser Enhanced Science - Department of Laser Application in Environmental Metrology Photochemistry and Agriculture

In the current study, the application of near-infrared diode laser irradiation as a new method for gene delivery in yeast has been studied and compared with chemical (LiAc) as a traditional method. The two treatments have been carried out on Saccharomyces cerevisiae (strain,NCYC1611a) with pBS4 plasmid DNA carrying hygromycin B resistance (hphr) andcyano-fluorescent protein (CFP) genes. This study aimed to establish a novel transformation method using 808nm near IR diode laser irradiation to facilitate introducing exogenous reporter genes in yeast. Results showed slight differences between the two method.NIR diode laser irradiation methods proved the best vs. LiAc chemical method because it showed less ultra- damaging and less cytotoxic effects compared to the control normal yeast.Histological and molecular analysis proved the presence, and expression of both hphr and CFP genes in the two methods. Molecular laser Raman Spectroscopy (MLRS) confirmed the presence of the pBS4 plasmid, containing both trans-genes in transformed yeast by the two methods. Also confocal laser scanning microscopy (CLSM) proved the expression of CFP gene. Ultra- damages was detected using transmission electron microscopy (TEM), Raman spectroscopy was usedalso to detect the damages on DNAmolecular scale for both methods vs. normal yeast cells

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