Cloning and prokaryotic expression of d-EGF and -defensin-3 fusion protein

To recombine the dGF and(3lefensin 3 gene in prokaryotic expression vector express fusion protein inProkaryotic system and purify and identify its specific. Methods:On the basis of gene sequence(3lefensin 3 gene and epidermalgrowth factor(EGF)gene were amplified by RTCR. Recombinant plasmid ofplefensin 3 and dGF plasmid were constructed andthe DNA sequences of dGF andplefensin 3 gene was cloned into the pET SUMO prokaryotic expression vector. The pET SUMOd-EGF vector was identified by PCR and sequenced and transformed into E. coli BL21(DE3),which was induced by IPTG purified usingNIrap column and identified with westernlot. Results:plefensin 3 and dGF plasmid were constructed. The fragment of 294 bywas successfully amplified by the recombinant PCR in accordance with the expected results and sequencing was correct. TwentyightkDa protein was obtained through tranforming into E. coli BL21 DE3)and IPTG Inducing. The fusion protein containing dGF andp-defensin 3 two proteins were identified with westernlot. Conclusions:Prokaryotic expression vector pET SUMOGF was successfully. onstructed and fusion protein purified was obtained. It provides a basis for analysing its activity