PRODUCTION OF INSULIN-FUSION PROTEIN USING GENETIC ENGINEERING

摘要

The present invention relates to a method for increasing the stability and expression rate of proinsulin fusion proteins in producing human proinsulin fusion proteins using genetic engineering.;First, the human proinsulin gene was cloned into E. coli host by cDNA cloning method. The cloned human proinsulin gene was introduced into pUC8 with the Lac ribosomal binding site gene, (Thr) 6 encoding [pUC8- (Thr) _6- PI], and pYK10 from pUC8- (Thr) _6- PI and pEX2. An -9 expression vector was prepared.;The fusion protein expressed in Escherichia coli in which pYK10-9 Berger with a short prosulin sequence was introduced was stable in E. coli cells. Escherichia coli pop2136 transformed with pYK10-9 expression vector produced 249 mg / l of human proinsulin fusion protein in flask fermentation, and the amount of proinsulin produced by fermentation of the bacterium was significantly increased to 2.2 g / l. .