Cloning and Expression of a Fused Gene Encoding TNFRI Death Domain and Chloramphenicol Acetyltransferase.
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.
Lin
Li-Zhu
LZ
Institute of Genetics, Fudan University, State Key Laboratory of Genetic Engineering, Shanghai 200433, China. shouyuanzhao@fudan.edu.cn
Li
Chang-Ben
CB
Zhao
Shou-Yuan
SY
De Wilde
G.
G
Haegeman
G.
G
Fiers
W.
W