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Molecular cloning and characterization of a glutathione S-transferase in the tropical liver fluke, Fasciola gigantica

Published online by Cambridge University Press:  21 July 2009

A. Jedeppa ,
O.K. Raina ,
S. Samanta ,
G. Nagar ,
N. Kumar ,
A. Varghese ,
S.C. Gupta  and
P.S. Banerjee
A. Jedeppa
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
O.K. Raina
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
S. Samanta*
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
G. Nagar
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
N. Kumar
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
A. Varghese
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
S.C. Gupta
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
P.S. Banerjee
Affiliation:
Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
*
*E-mail: [email protected]
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Abstract

Glutathione S-transferase from an Indian isolate of Fasciola gigantica of buffalo origin was isolated and characterized. Total RNA was transcribed to cDNA by reverse transcription and an amplicon of 657 bp glutathione S-transferase gene was obtained by polymerase chain reaction (PCR). The present isolate showed 99.1% sequence homology with the published sequence of the F. giganticaGST gene of cattle origin, with six nucleotide changes causing an overall change of four amino acids. Glutathione S-transferase protein was expressed in Escherichia coli using a prokaryotic expression vector pPROEXHTb. The recombinant protein was purified under non-denaturing and denaturing conditions by nickel nitrilotriacetic acid (Ni-NTA) affinity chromatography. Recombinant GST protein detected F. gigantica infection in naturally infected buffaloes by dot-ELISA.

Type
Research Papers
Information
Journal of Helminthology , Volume 84 , Issue 1 , March 2010 , pp. 55 - 60
Copyright
Copyright © Cambridge University Press 2009

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