Expression Of Soluble Insulin Glargine And Correction Of Genetic Diabetic Obese Mice

In this study, we aimed to provide innovative solutions for diabetes treatment from two aspects. First, we tried to achieve soluble expression of insulin glargine. Second, we worked on the correction of genetic diabetic obese mice.一、Expression of Soluble Insulin GlargineInsulin glargine is a therapeutic drug for diabetes and has an annual sale of more than three billion dollars. The quality and price of insulin glargine products are determined by the genetic engineering expression process. This study aimed to achieve soluble expression of insulin glargine. In our study, Trx (thioredoxin) was inserted between A chain and B chain of insulin glargine(ING) to achieve the soluble expression of insulin glargine. Soluble ING-Trx accounted for about 35% of total ING-Trx expression. Co-expression of molecular chaperones pGTF2 (GroEL、GroES、 TF) was also used to promote exact folding of ING-Trx. The results indicated that the production of soluble ING-Trx increased to 78% of total ING-Trx expression after co-expression with molecular chaperones. The production was about 4.5 mg/L after preliminary purification with three-fold more than that without molecular chaperones. This study may be a valuable exploration for genetic engineering production of soluble insulin glargine.二、Correction of Genetic Diabetic Obese Micedb/db mouse is a typical diabetic animal model with a mutation in leptin receptor gene. In this study, we used the db/db mice as a animal model to study correction of genetic diabetes. As a death signal adaptor protein in FAS/FasL apoptosis signal transduction pathways, FADD plays an important role in apoptosis. Previous studies in our laboratory revealed that FADD may also take part in metabolism. In the study, we mated FADD +/- TgD mouse with DB/db mouse to finally obtain db/db and FADD -/- TgD double mutant mouse. The results indicated that no embryonic lethal or aging phenomena were observed in db/db and FADD -/- TgD mouse, and obesity symptoms of db/db mouse were obviously reversed. However, obesity symptoms of db/db and FADD +/- TgD mouse were not changed, indicating that wild-type FADD will interfere with the obesity correction function of phosphorylated FADD on db/db mice.