Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
Vol. 10, No. 3, 2007, pp. 348-357
Bioline Code: ej07033
Full paper language: English
Document type: Research Article
Document available free of charge
Electronic Journal of Biotechnology, Vol. 10, No. 3, 2007, pp. 348-357
© Copyright 2007 - Pontificia Universidad Católica de Valparaíso -- Chile
A most effective method for selecting a broad range of short and medium-chain-length polyhidroxyalcanoate producing microorganisms|
Romo, Dolly Margot Revelo; Grosso, María Victoria; Solano, Nubia Carmenza Moreno & Castaño, Dolly Montoya
A molecular approach was used for selecting polyhydroxyalcanoate (PHA)-accumulating potential Gram-negative bacteria from different genera by colony polymerase chain reaction (PCR). Three degenerate primers were designed for amplifying a fragment from PHA synthase gene (phaC) (Class I), phaC1 and phaC2 (Class II) genes for detecting PHA-producing bacteria. Thirty-four out of 55 bacterial strains from the old collection selected using Sudan black B staining were phaC+. PCR was used for directly selecting 35 new collection bacterial strains; these strains were phaC+ and their ability to produce PHA was confirmed by Sudan black B staining. Four specific primers were designed on genes of Class II PHA biosynthesis operon. These primers were used for evaluating 9 strains from the old phaC+ collection; 6 showed Class II PHA synthase organisation. 34 from the old and new bacterial isolation were characterised by 16S ribosomal gene (16S rDNA) gene partial sequencing. The tool proposed here can be used for better directing PHA production based on PHA biosynthesis genes and bacterial genera. Class I or II phaC genes were detected in 9 different genera and were able to infer the type of polymer produced.
16S rDNA, colony PCR, degenerate primers, PHA synthase, polyhydroxyalcanoate, seminested PCR.
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