STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

• Main Author: T. VINTILĂ
• Format: Article
• Language: English
• Published: Agroprint Timisoara 2007-05-01
• Series: Scientific Papers Animal Science and Biotechnologies
• Subjects: genetic transformation; bacillus licheniformis; electroporation; protoplast; plc1; pnc61
• Online Access: http://spasb.ro/index.php/spasb/article/view/1535

A strain of Bacillus licheniformis was subject to genetic transformation with plasmid vectors (pLC1 and pNC61), using electroporation technique, protoplast transformation and bivalent cations (CaCl2) mediated transformation. In the case of transformation by electroporation of Bacillus licheniformis B40, the highest number of transformed colonies (3) were obtained only after a 1,79 KV electric shock, for 2,2 milliseconds. Using this transformation technique we have obtained six kanamycin resistant transformants. The frequency of Bacillus licheniformis B40 protoplasts transformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF = 10%). As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts, six kanamycin resistant transformants were obtained. The pNC61 plasmid, which confers trimethoprim resistance, does not integrate in receiver cells by protoplast transformation. The direct genetic transformation in the presence of bivalent cations (CaCl2), mediated by pLC1 and pNC61 plasmid vectors, produce a low transformation frequency. Using this technique, we have obtained three trimethoprim resistant colonies and four kanamycin resistant colonies. The chemical way of transformation is the only technique, which realizes the integration of pNC61 in B. licheniformis B40 cells.