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Title: Prokaryotic expression of chicken infectious anemia apoptin protein and characterization of its polyclonal antibodies
Authors: Saxena, Shikha
Kumar, Gandham Ravi
Singh, Prafull
Chaturvedi, Uttara
Saxena, Lovleen
Kumar, Rajiv
Sahoo, Aditya Prasad
Doley, Juwar
Kumar, Amit
Kumar, Sudesh
Tiwari, Ashok K
Keywords: Chicken infectious anemia;Polyclonal sera;Prokaryotic expression;VP3 gene
Issue Date: May-2012
Publisher: NISCAIR-CSIR, India
Abstract: In the present study recombinant VP3 (rVP3) was expressed in E.coli BL21 (DE3) (pLysS) and its polyclonal antibodies were characterized. SDS–PAGE analysis revealed that the expression of recombinant protein was maximum when induced with 1.5 mM IPTG for 6 h at 37ºC. The 6×His-tagged fusion protein was purified on Ni-NTA and confirmed by Western blot using CAV specific antiserum. Rabbits were immunized with purified rVP3 to raise anti-VP3 polyclonal antibodies. Polyclonal serum was tested for specificity and used for confirming expression of VP3 in HeLa cells transfected with pcDNA.cav.vp3 by indirect fluorescent antibody test (IFAT), flow cytometry and Western blot. Available purified rVP3 and polyclonal antibodies against VP3 may be useful to understand its functions which may lead to application of VP3 in cancer therapeutics.
Page(s): 325-331
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.50(05) [May 2012]

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