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Prokaruotic Expression of RhoA Gene and Its Tissue Location in Planarian Dugesia japonica
Nie Min, Zhao Bosheng*
Laboratory of Developmental and Evolutionary Biology, School of Life Sciences,Shandong University of Technology, Zibo 255049, China
Abstract: To investigate the certain function of RhoA in planarian Dugesia japonica， the DjRhoA gene was cloned into prokaryotic expression vector pBV220-IL1 using the methods of PCR， restriction digest and sequencing. The recombinant vector was then transferred into E.coli DH5α. The fusion protein RhoA expression was induced by 42 ºC and purified to detect with SDS-PAGE. The results indicated that the DjRhoA gene expressed as inclusion bodies in E.coli DH5α， and its molecular weight was estimated to be approximately 18 kDa. Western blot showed that the antibody reacted with the protein. Based on immunofluorescence analyses， RhoA was mainly expressed in the nervous system of planarian.
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