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 -