TY  - JOUR
T1  - Expression and One Step Purification of The Full-length 
Biologically Active, Nsp4 Of Human Rotavirus Wa Strain
AU - , Z. Sharifi AU - , B. yakhchali AU - , M.S. Shahrabadi 
JO  - International Journal of Molecular Medicine and Advance Sciences
VL  - 1
IS  - 3
SP  - 206
EP  - 212
PY  - 2005
DA  - 2001/08/19
SN  - 1813-176x
DO  - ijmmas.2005.206.212
UR  - https://makhillpublications.co/view-article.php?doi=ijmmas.2005.206.212
KW  - Expression
KW  -NSP4
KW  -rotavirus
KW  - E. coli
AB  - Rotavirus nonstructural glycoprotein, NSP4 has been proposed as the first viral enterotoxin capable of inducing diarrhea and a target for vaccine development. In order to study biological role of NSP4, a cDNA from human rotavirus (Wa strain) RNA segment 10 was amplified by PCR, using two specific and cloned into cloning vector pBS-KS(+) and subsequently into pQE-30 expression plasmid. Expression of NSP4 was demonstrated by SDS-PAGE,Western blot and ELISA using polyclonal antibody against NSP4 from SA11 infected BSC1 cells. The recombinant protein was purified by an affinity chromatography on Nickle NTA-Agarose column (Qiagen). A polyclonal antiserum against purified recombinant NSP4 was raised in Rabbit; which was reacted with NSP4 in BSC1 cells infected with SA11 rotavirus. Intraperitoneally inoculation of NSP4 caused diarrhea in BALB/c suckling mice indicating its biological activity. Intraperitoneal and oral inoculation of NSP4 antiserum significantly decreased diarrhea disease. These results indicated successful expression and purification of the full-length biologically active, NSP4 of human rotavirus Wa strain in E. coli and showed that antibody against it was able to protect against simian rotavirus diarrhea in neonatal mice.
ER  -