African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12227

Full Length Research Paper

Isolation, identification and PCR amplification of merA gene from highly mercury polluted Yamuna river

Md. Zeyaullah1, Badrul Islam2 and Arif Ali3*
1Faculty of Medicine, Omar Al-Mukhtar University, Al-Baida, Libya. 2Faculty of Pharmacy, Omar Al-Mukhtar University, Al-Baida, Libya. 3Gene Expression Laboratory, Department of Biotechnology, Jamia Millia Islamia, New Delhi- 110025, India.
Email: [email protected]

  •  Accepted: 20 July 2009
  •  Published: 14 June 2010

Abstract

 Mercury resistant Escherichia coli strains have been isolated from different mercury polluted sites of India and their minimum inhibitory concentration (MIC) levels were determined for HgCl2. Biochemical identification of E. coli strains was also performed with HiMedia’s biochemical identification kit for various biochemical assays, like, Citrate utilization, Lysine decarboxylase, Ornithine decarboxylase, Urease, Phenylalanine deamination, Nitrate reduction, H2S production, Glucose, Adonitol, Lactose, Arabinose, Sorbitol etc. The zone of inhibition was measured to find the antibiotic susceptibility level. The location of mer operon [containing mergenes including merA responsible for the expression of mercuric reductase enzyme (MerA) which converts the toxic Hg2+ to least toxic elemental Hg0] was determined by transforming the isolated plasmids into mercury sensitive host E. coliDH5α cells. Plasmid isolated from transformed E. coli DH5α cells were also analyzed and compared with the plasmid profile of the wild-type E. coli strains. Oligonucleotides primer were designed by comparing the known reported sequences of merA from Gram-negative bacteria (Escherichia coli plasmid R100) and 1695 bp of merA gene was amplified by PCR.

 

Key words: Mercury resistant, Escherichia coli, minimum inhibitory concentration (MIC), mer operon, merA.