@article{MAKHILLIJMMAS2005138495,
    title = {Expression and One Step Purification of The Full-length 
Biologically Active, Nsp4 Of Human Rotavirus Wa Strain},
    journal = {International Journal of Molecular Medicine and Advance Sciences},
    volume = {1},
    number = {3},
    pages = {206-212},
    year = {2005},
    issn = {1813-176x},
    doi = {ijmmas.2005.206.212},
    url = {https://makhillpublications.co/view-article.php?issn=1813-176x&doi=ijmmas.2005.206.212},
    author = {Z. Sharifi,B. yakhchali and},
    keywords = {Expression,NSP4,rotavirus, E. coli},
    abstract = {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.}
    }