en Biological Sciences Biological Sciences Original Research: Basic Science in Medicine Original Research: Basic Science in Medicine Rotavirus nonstructural glycoprotein NSP4 can induce diarrhea in newborn mice. It has been suggested that NSP4 may be a key determinant for rotavirus pathogenesis and a target for vaccine development. In order to study the biological and morphological role of NSP4 a large amount of the purified protein and antibody against it are required. Simian rotavirus SA11 was propagated in BSCl cell, purified on cesium chloride gradients, and its genomic RNA was extracted. A cDNA from RNA segment 10 was synthesized and amplified by RT-PCR. cDNA fragment was cloned into plasmid vectors. The recombinant plasmid was characterized by restriction enzyme and sequencing. Construction of expression plasmid containing nsp4 gene was performed and expression of NSP4 was demonstrated by SDSPAGE, Western blot, ELISA and IF using polyclonal antibody against NSP4 from SA11 infected BSCI cells. A polyclonal antiserum against purified recombinant NSP4 was raised in rabbit which was reacted with NSP4 in BSCI cells infected with SA11 rotavirus. These results indicated successful expression of the full-length NSP4 in E.coli to produce antibody against it and to study its biological activities. Expression, NSP4, Rotavirus, E.coli. 217 224 http://mjiri.iums.ac.ir/browse.php?a_code=A-10-298-144&slc_lang=en&sid=1 Z SHARIFI 20031947532846003738 20031947532846003738 No 123From the Virology Department, Iran University of Medical Sciences, Tehran, the National Research Center for Genetic Engineering and Biotechnology (NRCGEB), Tehran, and the Iranian Blood Transfusion Organization, Tehran, Iran, B YAKHCHALI byakhcha@nrcgeb.ac.ir 20031947532846003739 20031947532846003739 Yes National Research Center for Genetic Engineering and Biotechnology (NRCGEB), Tehran MS SHAHRABADI 20031947532846003740 20031947532846003740 No From the Virology Department, Iran University of Medical Sciences, Tehran