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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.50 [2012] >
IJEB Vol.50(05) [May 2012] >
| 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
Rajmani
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 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.50(05) [May 2012]
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