Cloning And Expression The Fragment Of Tudor Of Human SMN Sequence

Objective: Constructing the expression vector containing the sequence of SMN Tudor, in order to compare the function of SMN Tudor with p100 Tudor.Methods: DNA fragments corresponding to Tudor of human SMN was generated by PCR amplification. The fragment were inserted into the vector pGEX-4T-1 and then transformed to E. coli strain DH5a. After identified by limited cutting enzyme digesting and sequenced the vector transformed into E. coli strain BL21 again. The expression of GST SMN Tudor fusion protein was induced by IPTG and then purified by using glutathione-Sepharose.Results: The fragment of SMN Tudor produced by PCR were 183bp in size, and the protein of SMN Tudor was expressed successfully in E. coli strain BL21. Forethemore, the nuclear proteins which bind to SMN Tudor are different from the ones bind to p100 Tudor.Conclusion: The cloning of SMN Tudor gene and the construction of the expression vector pGEX-4T-1- SMN Tudor are achieved.