원문정보
초록
영어
Overexpression of cytoplasmic transduction peptide (CTP: YGRRARRRRRR) conjugated recombinant interferon-α-2b (CTP-IFN) was achieved by synthesizing codon optimized gene and expressing as inclusion bodies (IBs) in Escherichia coli. To overcome the high cost and the difficulties in optimizing expression of IPTG induction system, we introduced the CTP-IFN gene into E. coli pop2136 by ligation with pC (plasmid of Creagene) vector, which contains heat inducible λPL promoter. Expression of CTP-IFN was optimized with considering the media, induction initiation point and induction time. The Volumetric yields of CTP-IFN reached 30g/L at final OD600 of 28 with 5L bioreactor. The solubilized IB in 8M urea as a denaturant was refolded with drop-wise renaturation process. The refolded proteins were applied to cation exchange
chromatography and gel filtration chromatography and finally, we got highly pure CTP-IFN (purity more than 98%).