Volume No. 4 Issue No.: 1 Page No.: 232-238 July-Sept 2009

 

PCR AMPLIFICATION OF dmpL GENE IN METAGENOMIC D.N.A. USING ACCLIMATIZATION-ENRICHMENT METHOD

 

Veena Dhanwani*, Rita Tonge, S.D. Bagchi, Meenakshi Mundafode,
Damini Motwani, M.L. Jisnani an

SMV Centre for Biotechnology, Sindhu Mahavidyalaya,
Panchpaoli, Nagpur-17, Maharashtra (INDIA)

 

Received on : March 30, 2009

 

ABSTRACT

 

Environmental pollutants like xenobiotic compounds released from industrial effluents and domestic wastes are major threat to mankind. These compounds lead to bioaccumulation in the environment. Nitroaromatic compounds (NACs) and their derivatives used in various industries, play a major role in contributing xenobiotic pollution in nature. On entering mammalian system, NACs are readily reduced to more reactive and potentially more carcinogenic or mutagenic derivatives leading to many health related problems. Our present work is aimed at studying such micro-organisms involved in biodegradation of 4-nitrophenol, a major xenobiotic pollutant. Biodegradation of xenobiotics by microbial enzymes are often produced by induction process, hence we enriched the soil samples collected from Godavari river at Nashik with varying concentrations from 10-70mM of 4-nitrophenol. The OD of these cultures of varying concentration of 4-nitrophenol was measured at 620nm after every 6 hrs interval. It was found that number as well as types of the organisms decreased with increased concentration of 4-nitrophenol. The enriched Metagenomic DNA of all the culture samples was isolated and PCR amplification of dmpL gene was carried out. The dmpL gene is involved in phenol degradation. The amplicons obtained from all the samples indicated the presence of phenol hydroxylase gene. The amplicons are to be sent for sequencing for its further study. The work is being further extended in view of isolation, identification and characterization of the isolated species and other genes involved in phenol biodegradation.

 

Keywords : Xenobiotic, Phenol biodegradation, 4-nitrophenol, Phenol hydroxylase, dmpL gene

 

 

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