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Cloning, expression and sequence analysis of Macrobrachium rosenbergii nodavirus genes: Indian isolate
Author(s) -
Shekhar Mudagandur S,
Sahoo Pramoda K,
Dillikumar Manickam,
Das Abhilipsa
Publication year - 2011
Publication title -
aquaculture research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 89
eISSN - 1365-2109
pISSN - 1355-557X
DOI - 10.1111/j.1365-2109.2010.02775.x
Subject(s) - biology , capsid , microbiology and biotechnology , virology , sequence analysis , gene , recombinant dna , virus , genetics
RNA‐dependent RNA polymerase (RdRp), B2 and capsid genes of Macrobrachium rosenbergii nodavirus ( Mr NV) of Indian isolate were polymerase chain reaction amplified, cloned and sequenced. Expression of the Mr NV 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 Mr NV capsid protein showed specificity against the capsid protein by Western blot. The protein sequence analysis of the partial RdRp gene of Mr NV 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 Mr NV showed high RdRp and capsid gene sequence homology with the other Mr NV 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 Mr NV 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.