Protein Glycosylation Engineering

IL-2 gene cloning and expression in S,cerevisiae, EPO gene cloning and expression in E.coli,Yeast,and mammanlian cellsWith the enormous advances in our understanding of glycoproteins,It has been more atractive to engineering the oligosaccharides. Scientists began to enhance the therapeutic potential of recombinant proteins by the metheds of glycosylation engineering which are as followings: (1)product ion in a specific host cells and corresponding glycosylation mutants,(2) mutation of the sites of glycosylation (3)Production in the presence of glycosylation inhibitors (4) treatment of purified glycoprotein with glycosidase or glycosyltransferases.We set about the study from gene cloning and expression on two typical glycoprotein: IL-2 and EPO which represent O-type and N-type respectively. The research divided into four parts:1. Interleukin- 2 gene cloning and expression in S. cerevisiaeMature human interleukin-2 gene was amplified from IL-2 cDNA PCR methods. The PCR product was cloned into pUC12 plasmid at SmaI site, then precisely engineered into a...