Construction and Expression of Recombinant Plasmids of Tudor-SN Protein TSN Domain’s Sub-Fragments

Objective: To Construct recombinant plasmids pEGFP-C2-Tudor-SN-TSN (Ⅰ-Ⅳ) for the further functional research of human Tudor-SN TSN domain’s 4 sub-fragments. 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 plasmid pEGFP-C2-TSN(Ⅰ-Ⅳ) was transfected into HeLa cells. The expression of fusion proteins were examined by fluorescent microscopy and Western blot. Results: The sub-fragments of TSN domain can be detected in products of the restriction double enzyme digestion. The green fluorescent fusion proteins were observed in HeLa cells after transfection. The fusion proteins pEGFP-C2-TSN(Ⅰ-Ⅳ) can be detected from the lysate of transfected HeLa cells by Western blot. Conclusion: These recombinant eukaryotic plasmids,pEGFP-C2-Tudor-SN-TSN(Ⅰ-Ⅳ),were constructed successfully and expressed effectively.