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Published on: June 25, 2015
Cloning and Expression of a Fused Gene Encoding TNFRI Death Domain and Chloramphenicol Acetyltransferase
Li-Zhu Lin1, Chang-Ben Li, Shou-Yuan Zhao
1Institute of Genetics, Fudan University, State Key Laboratory of Genetic Engineering, Shanghai 200433, China. shouyuanzhao@fudan.edu.cn
Abstract:
By designing four primers for an overlapping PCR, we created a fusion gene Ddlcat encoding human TNF receptor I death domain and chloramphenicol acetyltransferase (CAT). By DNA sequencing, the whole sequence of the fusion gene is confirmed to be correct. Two hours after induction with IPTG, we could see a 39 kD extra protein band on SDS-PAGE pattern. We proved that this 39 kD protein band is DdLcat protein by Western blotting. Then we purified this protein to the purity of 95% through Q-Sepharose chromatography.
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