Expression of mouse tuftelin in E. coli and preparation of antiserum against purified tuftelin

AIM:To express tuftelin and purify it from E.coli and prepare its antiserum. METHODS :The plasmid pPRoEX TM HTc and the sequence of Tuftelin cDNA(365 amino acid)cloning in the plasmid pBluescript KS+ were cut by the restricted endonucleas.Then the two fragment were ligased by T4 DNA ligasing and the expression plasmid of fusion protein was constructed.The product was transformed into E.coli . After the induce of IPTG, the E.coli was centrifuged and breakaged by SDS loading buffer. The purified tuftelin was mixed with incomplete adjuvant, and then used to immunize rabbits. RESULTS:The product of M r 42×10 3 fusion protein was in accordance with the anticipation. 8 weeks later. the antiserum titer of 1:100 000 could be showed by the test of Western blot. CONCLUSION :The tuftelin is expressed successfully and the antiserum is prepared.