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

T. VINTILĂ

Abstract


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.


Keywords


genetic transformation; Bacillus licheniformis; electroporation; protoplast; pLC1; pNC61

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LUCRARI STIINTIFICE ZOOTEHNIE SI BIOTEHNOLOGII (SCIENTIFIC PAPERS ANIMAL SCIENCE AND BIOTECHNOLOGIES)

ISSN print 1841 - 9364
ISSN online 2344 - 4576
ISSN-L  1841 - 9364
(former ISSN 1221-5287, E-ISSN 1841-9364)

PUBLISHER: AGROPRINT Timisoara, Romania
PAPER ACCESS: Full text articles available for free
FREQUENCY: Semiannual
PUBLICATION LANGUAGE: English
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Banat´s University of Agricultural Sciences and Veterinary Medicine "King Michael I of Romania" from Timisoara
Bioengineering Faculty of Animal Resources
300645, Timişoara, Calea Aradului 119, Romania

E-mail: spasb [at] animalsci-tm.ro
Phone: +40-256-277160, Fax.: +40-256-277110