Cloning, expression and sequence analysis of Macrobrachium rosenbergii nodavirus genes: Indian isolate

  1. Mudagandur S Shekhar1,
  2. Pramoda K Sahoo2,
  3. Manickam Dillikumar1,
  4. Abhilipsa Das2

Article first published online: 25 JAN 2011

DOI: 10.1111/j.1365-2109.2010.02775.x

Issue

Aquaculture Research

Aquaculture Research

Volume 42, Issue 12, pages 1778–1788, November 2011

Additional Information

How to Cite

Shekhar, M. S., Sahoo, P. K., Dillikumar, M. and Das, A. (2011), Cloning, expression and sequence analysis of Macrobrachium rosenbergii nodavirus genes: Indian isolate. Aquaculture Research, 42: 1778–1788. doi: 10.1111/j.1365-2109.2010.02775.x

Author Information

  1. 1

    Genetics and Biotechnology Unit, Central Institute of Brackishwater Aquaculture, Chennai, India

  2. 2

    Central Institute of Freshwater Aquaculture, Fish Health Management Division, Bhubaneswar, India

*Correspondence: Dr M S Shekhar, Genetics and Biotechnology Unit, Central Institute of Brackishwater Aquaculture, 75 Santhome High Road, R. A. Puram, Chennai, India. E-mail: msshekhar@hotmail.com

Publication History

  1. Issue published online: 14 NOV 2011
  2. Article first published online: 25 JAN 2011

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Keywords:

  • Macrobrachium rosenbergii nodavirus;
  • RNA-dependent RNA polymerase (RdRp);
  • B2;
  • capsid;
  • recombinant proteins

Abstract

RNA-dependent RNA polymerase (RdRp), B2 and capsid genes of Macrobrachium rosenbergii nodavirus (MrNV) of Indian isolate were polymerase chain reaction amplified, cloned and sequenced. Expression of the MrNV fusion recombinant proteins of RdRp (44.5 kDa), B2 (32.2 kDa) and capsid (58.4 kDa) was confirmed by Western blot analysis using anti-His mouse monoclonal antibodies. Polyclonal antibodies specific to purified recombinant MrNV capsid protein showed specificity against the capsid protein by Western blot. The protein sequence analysis of the partial RdRp gene of MrNV revealed the signature sequence along with the conserved core residues of the catalytic domain and indicated the presence of active sites, metal ion-binding site and nucleic acid-binding site residues. The Indian isolate of MrNV showed high RdRp and capsid gene sequence homology with the other MrNV geographical isolates. However, the Belize isolate was found to be the most distinct among the different geographical prawn nodavirus isolates due to the host specificity. Secondary structure prediction analysis of the MrNV capsid predicted it to be a DNA-binding protein consisting of α helix (22.91%), extended strand (24.80%), β turn (5.39%) and random coil (46.90%) regions.

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