PRODUCTION AND PURIFICATION OF PLASMID VECTOR PUB110 IN TRANSFORMED BACILLUS SUBTILIS BACTERIA | ||
| Iraqi Journal of Biotechnology | ||
| Article 1, Volume 11, Issue 2, December 2012, Pages 141-150 | ||
| Author | ||
| Khlood A. Al- Khafaji | ||
| Abstract | ||
| Ninety percent of B. subtilis protoplast could obtained after 30 minute from treatment of the bacteria with both Ethylene diamine tetra acetic acid (EDTA) and lysozyme enzyme. Protoplast transformed with the plasmid vector pUB110 had efficiency 104 cell/mg of DNA . Kanamycin resistant cell was gained after culturing transformed bacteria onto regeneration medium which containing kanamycin antibiotic. Three different methods were used for DNA extraction from transformed B. subtilis and the plasmid vector pUB110 could detected from the three methods. Salting out procedure gave the highest DNA yield with chromosomal DNA smear in comparison with the other two procedures. Furthermore, gel electrophoresis for DNA extracted by CTAB method showed high purity DNA extract in comparison with other extraction methods. | ||
| Keywords | ||
| pUB; Bacillus subtilis; transformation; Protoplast | ||
|
Statistics Article View: 146 PDF Download: 62 |
||