IMPROVED TRANSFORMATION EFFICIENCY OF DEXTRAN GENE(S) IN Saccharomyces cerevisae USING THE COMBINED EFFECT OF TWO DIFFERENT DIRECT TRANSFORMATION METHODS | ||
| Journal of Agricultural Chemistry and Biotechnology | ||
| Article 1, Volume 25, Issue 3, March 2000, Pages 1797-1803 PDF (581.99 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/jacb.2000.258764 | ||
| Author | ||
| Fatma M. Hafez, | ||
| Microbial Genetics Department, Genetic Engineering and Biotechnology Div., National Research Center, Dokky - Cairo | ||
| Abstract | ||
| Intact yeast cells Saccharomyces cerevisiae can be efficiency transformed by combined effect of polyethylene glycol and CaCl2. This study had been undertaken to determine the effect of using 24% (PEG) in 0.4M manitol and 30 mM MgCl2, 0.3% CaCl2 and the combination of the two individuals. The results showed that (PEG) or CaCl2 alone caused a few transformants and a low of transformation percentage. In addition, (PEG) gave higher transformants and transformation percentages more than that of CaCl2 as it facilitates the entrance of DNA into the yeast cell walls. In contrast, the combined effect of (PEG) and CaCl2 in the same condition reproducibly yield the maximum number of transformants and transformation percentages. Data also revealed that the transformation percentage was 2.5 fold higher than values given in individual treatment. It is obvious that higher culture viscosity were produced by transformants isolates from combined effect and CaCl2 experiments. This observation was due to Ca++ activated dextran sucrose by increasing the Vmax and decrease the Km of sucrose polymerase. In this study, transformants isolates seem to been differ in their ability to express dextran gene(s). This means that the events at one molecule can affect event at a distant location by different ways on the same molecule. | ||
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