Abstract: Abstract Objective:To construct eukaryotic green fluorescent protein(GFP) expressing recombinant plasmids, pEGFP-C2-Tudor-SN-TSN(Ⅰ~Ⅳ), which contain 4 sub-fragments of human Tudor-SN TSN domain, respectively. Methods: The genes of Tudor-SN-TSN fragments were amplified by PCR from the recombinant pSG5-Tudor-SN-flag plasmid then the EcoRI/Sal I digested fragments were combined with pEGFP-C2 vector. The recombinant expression plasmids pEGFP-C2-TSN(Ⅰ~Ⅳ) were transfected into HeLa cells. The expression of fusion proteins were examined by fluorescent microscopy and Western blot. Results: (1) The sub-fragments of TSN domain can be detected in the products of the restriction double enzyme digestion; (2) The green fluorescent fusion proteins were observed in HeLa cells after transfection; (3) The fusion proteins pEGFP-C2-TSN(Ⅰ~Ⅳ) can be detected from the lysate of transfected HeLa cells by Western blot. Conclusion: (1) The expression plasmids pEGFP-C2-TSN(Ⅰ~Ⅳ) was successfully constructed. (2) These recombinant eukaryotic plasmids can be used for protein detection by binding with anti-GFP antibody for the further study on molecular mechanism in biologic function of Tudor-SN-TSN.

Key words: Human Tudor-SN protein, SND1, pEGFP-C2, Recombinant plasmid, Fusion protein