Several factors affecting the transformation of the yeast Saccharomyces cerevisiae by electroporation / by Lawrence S. Graham.
| Author/creator | Graham, Lawrence S. author. |
| Other author | Smith, W. James, degree supervisor. |
| Other author | East Carolina University. Department of Biology. |
| Format | Theses and dissertations |
| Production | 1990. |
| Description | [v]. 56 leaves : illustrations ; 28 cm |
| Supplemental Content | Access via ScholarShip |
| Subjects |
| Summary | Electroporation is a relatively new technique for the transformation of prokaryotic and eukaryotic cells with exogenous DNA. Although electroporation has been used with great success in mammalian cell studies, relatively few studies have investigated improving the technique in the transformation of yeast cells. We have investigated several factors which enhance the transformation of whole Saccharomyces cerevisiae cells and spheroplasts by capacitance-discharge electroporation. Factors tested include electroporation voltage and capacitance parameters, electroporation temperature, the presence or absence of carrier DNA, buffer pH, buffer salt composition, DNA concentration, cell culture stage, and treatment of whole cells with proteinase K prior to electroporation. From these experiments a protocol for the rapid transformation of yeast cells was developed. The protocol reliably produces 5-7 X 10³ transformants per microgram of YRpl7 DNA, and as many as 1.2 X 10⁴ per microgram have been recovered. The protocol requires about one hour excepting culture growth time and can be a useful asset for any studies requiring the transformation of yeast. |
| General note | Submitted to the faculty of the Department of Biology. |
| General note | Advisor: W. James Smith |
| Dissertation note | M.S. East Carolina University 1990 |
| Bibliography note | Includes bibliographical references (leaves 51-56). |
| Genre/form | Academic theses. |
| Genre/form | Academic theses. |
| Genre/form | Thèses et écrits académiques. |