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  • Title: [Fusion expression of gastric cancer-related gene MPS-1 and preparation of rabbit antibody against GST-MPS-1].
    Author: Wang YW, Zhu ZG, Gu QL, Li JF, Qu Y, Liu BY, Lin YZ.
    Journal: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi; 2006 Jan; 22(1):60-3. PubMed ID: 16388747.
    Abstract:
    AIM: To express the fusion protein GST-MPS-1 in E.coli and prepare rabbit antibody against GST-MPS-1. METHODS: MPS-1 cDNA was inserted into the vector of pGEX-5X. The recombinant was then transformed into E.coli BL21. After induction with the IPTG, the fusion protein GST-MPS-1 was expressed in E.coli. The purified fusion protein was then used to immunize the New Zealand rabbits to obtain the polyclonal antibody against GST-MPS-1. Specificity of the antibody was tested by Western blot and immunofluorescent staining. RESULTS: The cDNA sequence of MPS-1 in the recombinant was confirmed by restriction endonuclease digestion and sequence analysis. SDS-PAGE result showed that fusion protein was highly expressed in E.coli with a molecular weight of 36 kDa. The titer of the rabbit serum against GST-MPS-1 was as high as 1x10(5) in ELISA analysis. The polyclonal antibody were found to specifically bind to MPS-1 protein in Western blot and immunofluorescent staining. CONCLUSION: The preparation of the polyclonal antibody against MPS-1 lay a foundation for the further study of MPS-1 expression at protein levels and its function.
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