Exendin-4/Human IgG-Fc 융합유전자 클로닝 및 인슐린 분비 촉진연구 : Cloning of Exendin-4/Human IgG-Fc Fusion Plasmid and Studying of the effect on Insulin Secretion

다국어 초록 (Multilingual Abstract)

Exendin-4 is a hormone found in the saliva of the Gila monster. It displays biological properties similar to human glucagon-like peptide-1 (GLP-1), a regulator of glucose metabolism and insulin secretion. GLP-1 possesses several physiological properties that make it a subject of intensive investigation as a potential treatment of diabetes mellitus, Because exendin-4 evades the degradation by dipeptidyl peptidase (DPP-Ⅳ). exendin-4 has a much longer half life then GLP-1; it is metabolized in vivo with half-life of 18–41 minutes.

In this study we have cloned exendin-4/human IgG-Fc fusion plasmid and expressed the fusion protein of exendin-4/human IgG-Fc in E. coli to seek its therapeutic possibility for the type 2 diabetes mellitus. To elucidate whether the exendin-4/human IgG-Fc fusion protein has retained bioactivity, we treated the rat β-cell line INS-1 cells with either GLP-1 or purified exendin-4/human IgG-Fc. Treatment of GLP-1 and exendin-4/human IgG-Fc both increased the expression of IRS-2. These results indicate exendin-4/human IgG-Fc might have possibly benefiting clinical usage. The effect of Fc fusion on prolonging the half-life of exendin-4 needs to be further determined.

목차 (Table of Contents)

• Ⅰ. Introduction (1)
• 1. Type 2 diabetes mellitus (1)
• 2. Insulin (2)
• 3. Glucagon-like peptide-1 (GLP-1) (5)
• 4. Exendin-4 (7)
• 5. Objective (9)
• Ⅱ. Materials and Methods (11)
• 1. Materials (11)
• 2. Preparation of exendin-4/human IgG-Fc Fusion Plasmid (12)
• 3. Gene expression of exendin-4/human IgG Fc region on pDHF28 vector (16)
• 4. Separation and purification of exendin-4/human IgG-Fc protein (17)
• 5. Treatment of exendin-4/human IgG-Fc protein into INS-1 Cells (18)
• Ⅲ. Results (20)
• 1. Preparation of exendin-4/human IgG-Fc Fusion Plasmid(20)
• 2. Gene expression of exendin-4/human IgG Fc region on pDHF24 vector 29)
• 3. Separation and purification of exendin-4/human IgG-Fc protein (29)
• 4. Treatment of exendin-4/human IgG-Fc protein into INS-1 Cells (36)
• Ⅳ. Discussion (37)
• Ⅴ. Reference (38)