We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Cloning, expression of the major capsid protein gene from marine algae Emiliania huxleyi virus and the possible use in detection of virus infection.
- Authors
Jingwen Liu; Zhilan Zhang; Xuhong Liu; Yiqin Cai; Huinong Cai
- Abstract
Here we described the cloning, bioinformatic characterization and expression in Escherichia coli of the major capsid protein (MCP) from marine unicellular algae Emiliania huxleyi virus EhV-99B1 isolate. The purified recombinant MCP was used to develop a polyclonal antibody for testing viral infection. The full length openreading frame (ORF) of MCP encodes a protein of 496 amino acids with a calculated molecular mass of 55 kDa and Ip 6.34. Hydropathy analysis of MCP showed that there were 6 largely hydrophobic domains, which may be important for the interaction with the envelope protein. The conserved region of EhV strains MCP had high similarity in amino acid sequence and secondary structure which allow us to develop a specific biomarker for EhVs infection detection. The full length ORF was subcloned into expression vector pGEX-4T-3 for overexpression in E. coli as glutathione-Stransferase-L1 (GST-L1) fusion protein and the soluble recombinant protein was used to generate polyclonal antibodies in mice. The obtained antisera reacted in Western immunoblots with the same protein both in purified EhV-99B1 virions and infected host cells sample. These shows that the antiserum against recombinant EhV-MCP offers the potential to develop immunofluorescence techniques for the detection of EhVs infected cells.
- Subjects
MARINE algae; COCCOLITHUS huxleyi; GENETIC engineering; VIRUS diseases; ESCHERICHIA coli; HYDROTHERAPY; RECOMBINANT proteins
- Publication
Microbiology Research, 2013, Vol 4, Issue 1, p21
- ISSN
2036-7481
- Publication type
Article
- DOI
10.4081/mr.2013.e5