Proposal of a method for the genetic transformation of Gordonia jacobaea.
OBJECTIVE
Gordonia jacobaea is a recently isolated bacterial species with potential industrial application on account of its ability to store large quantities of trans-canthaxanthin. Its genetic manipulation is, however, difficult and cumbersome owing to the presence of mycolic acids in the cell wall and, especially, because of current lack of knowledge about its basic genetics. The present work describes a method for the genetic transformation of G. jacobaea.


RESULTS
Gordonia jacobaea was grown in media supplemented with different glycine, penicillin G and isoniazid concentrations. The temperature, carbon source, growth phase and ultrasounds were analyzed for improving the method efficiency. The cells were finally transformed by electroporation. Finally, the method was applied to Brevibacteriumlactofermentum and Gordonia bronchialis.


CONCLUSIONS
The growth of G. jacobaea in the presence of glycine and isoniazid is essential for obtaining electrocompetents cells. The temperature, growth phase and ultrasounds appeared as the main factors for increasing the transformation efficiency. The use of shuttle plasmids became necessary. The method described can be used with other Corynebacteria species.


CONCLUSIONS
Because of the importance of the CNM group (Corynebacteria, Nocardia and Mycobacteria genera) in different areas such as industry, bioremediation improve the knowledge of their molecular mechanisms are becoming essential. The method described here improves the genetic manipulation of this group of bacteria.
Department of Microbiology, Faculty of Pharmacy, University of Santiago de Compostela, A Coruña, Spain.
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Veiga-Crespo
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Feijoo-Siota
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de Miguel
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Poza
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TG
Villa
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001951:Brevibacterium / Q000235:genetics; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D018274:Electroporation / Q000379:methods; D005998:Glycine / Q000494:pharmacology; D018684:Glycine Agents / Q000494:pharmacology; D033601:Gordonia Bacterium / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D007538:Isoniazid / Q000494:pharmacology; D010400:Penicillin G / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping / Q000379:methods; D014465:Ultrasonics
D000900:Anti-Bacterial Agents; D003470:Culture Media; D004269:DNA, Bacterial; D018684:Glycine Agents; D010400:Penicillin G; D005998:Glycine; D007538:Isoniazid