International Journal of
Biotechnology and Molecular Biology Research

  • Abbreviation: Int. J. Biotechnol. Mol. Biol. Res.
  • Language: English
  • ISSN: 2141-2154
  • DOI: 10.5897/IJBMBR
  • Start Year: 2010
  • Published Articles: 100

Full Length Research Paper

Molecular characterization of a gene capable of degrading trichloroethylene, an environmental pollutant

Srijata Mitra and Pranab Roy*
Department of Biotechnology, The University of Burdwan Golapbag Burdwan, West Bengal, India.
Email: [email protected]

  •  Accepted: 17 July 2011
  •  Published: 30 September 2011

Abstract

Trichloroethylene (TCE) is a widely used organic solvent and metal degreasing agent, one of the most frequently detected groundwater contaminants and a potential health hazard. Our novel isolate, Bacillus cereus strain 2479 was capable of degrading TCE efficiently. The gene for TCE degradation was PCR amplified from genomic DNA of B. cereus 2479. The amplified gene was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The sequencing results showed that this novel gene (designated as tce1, GenBank Accession No: GU183105) contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide is 5.17'. E. coli expressing the tce1 gene overproduces a polypeptide in the presence of the inducer Isopropyl-β-D-thiogalaocto-pyranoside which reacts immunologically to the polyclonal antibody against TCE inducible proteins of the strain 2479. The secondary structure of Tce1 protein was predicted through internet resources with software, CLC Protein Workbench. The present study suggested that cloned gene product (Tce1) was capable of degrading TCE as verified chemically.

Key words: Trichloroethylene, tce1gene, molecular cloning, expression, bioinformatics analysis.