Production of polyclonal antiserum against recombinant VP28 protein and its application for the detection of white spot syndrome virus in crustaceans.
The VP28 gene of white spot syndrome virus (WSSV) was cloned into pRSET B expression vector. The VP28 protein was expressed as a protein with a 6-histidine taq in Escherichia coli GJ1158 with NaCl induction. Antiserum was raised against this recombinant-VP28 protein in rabbits and it recognized VP28 protein in naturally and experimentally WSSV-infected shrimp, marine crabs, freshwater prawns and freshwater crabs. The antiserum did not recognize any of the other known WSSV structural proteins. Various organs such as eyestalks, head muscle, gill tissue, heart tissue, haemolymph, tail tissue and appendages were found to be good materials for detection of WSSV using the antiserum and detection of WSSV was successful in experimentally infected Penaeus monodon and P. indicus at 12 and 24 h post-infection (p.i.), respectively. The antiserum was capable of detecting WSSV in 5 ng of total haemolymph protein from WSSV-infected shrimp.
Yoganandhan
K
K
Aquaculture Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam, Tamil Nadu, India.
Syed Musthaq
S
S
Narayanan
R B
RB
Sahul Hameed
A S
AS
D000818:Animals; D017756:Aquaculture; D015153:Blotting, Western; D017931:DNA Primers; D004267:DNA Viruses / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli; D006458:Hemolymph / Q000821:virology; D007106:Immune Sera / Q000276:immunology*; D033561:Penaeidae / Q000821:virology*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D012965:Sodium Chloride; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D017931:DNA Primers; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D012965:Sodium Chloride