Cloning and Expression of an Organophosphate Degrading Gene (opdK | 24889
Journal of Proteomics & Bioinformatics

Journal of Proteomics & Bioinformatics
Open Access

ISSN: 0974-276X

Cloning and Expression of an Organophosphate Degrading Gene (opdK) from a Novel Bacterium Kocuria sp

N. Nagavardhanam and Z. Vishnuvardhan


Abstract :

Eight bacterial isolates (Y1-Y8) capable of degrading organophosphate pesticide were screened from contaminated agricultural soils. Isolate Y4 is found to be very effective in hydrolyzing (800mg/L) chlorpyrifos in M9 mineral medium. Morphological, cultural and biochemical analysis in combination with molecular characterization using PCR amplification of 16S rRNA of Y4 isolate revealed that it belongs to genus Kocuria showing 65% similarity with K.rosea. The report forms the first evidence that Kocuria degrades chlorpyrifos. The organophosphate hydrolyzing gene (named opdK) was amplified by using specific primers. It was further purified and ligated into pre-digested PUC19 vector. The ligated product was transferred into E.coli DH5α. The coding region of opdK gene was sub cloned into vector pRSETA and over expressed in E.coli BL 21. The study revealed that the cloned opdK gene was very effective in expression in E.coli BL 21.