Blood Glucose Regulation Related Genes And Functional Studies

Diabetes is a disease difficult to treat in worldwide. It is the third disease followed cancer and cardiac disease and one of five diseases with the highest incidence and mortality. If we don't try to treat it, in the following thirty years, diabetes sufferer in China will reach 5.0-10.0% which mean the total patient number will reach one hundred million. It is necessary to do research on etiology, underlying basis and best precautionary method of diabetes as soon as possible. Diabetes is a multigenes related disease with obvious inherited tendency which has extremely complex basis. In clinical, diabetes is divided into two group: type I diabetes (IDDM) and type II diabetes (NIDDM), the latter is 90% of total diabetes patient. There are two main strategies in NIDDM gene identification: candidate gene strategy and whole genome scanning strategy. Candidate gene strategy is to select genes participating in glucose metabolism and signal transduction as candidate genes. This strategy is simple and one of main strategy in NIDDM research now. But, as yet the result discovered by this strategy is less. This leads to speculate that NIDDM genes are possible those genes have not been discovered. For this purpose, it is necessary to discover glucose metabolism related new genes with proper methods.We have provided a new thought in the research of diabetes here: at first to cloning glucose metabolism related gene, secondly to discover how the related new gene participating into glucose metabolism, to provide basis of identify new candidate gene and of exploit new precautionary method.Firstly, we established SD rat glucose auto-regulating model by use of Jugular Vein right atrium intubation. This model is easy to make and has good repetition, has less artificial interference. Then mRNA differential display assay had been done by the use of this model.mRNA differential display is a method to isolate differential expressed genes from mammalian cell. There has been many modifications since this method has been established which made it a mature method. These modifications include: the number of anchor primers can reduce from 12 to 4; longer primers are used to make PCR more stringent; two-step PCR can enhance reliability. Beside we adopted these modifications, we adopted a strategy that first screening hundreds of differentially expressed bands through slot blot, then cloning and sequencing candidate cDNAs, Northern Blot ensure the differentially expressed genes at last. Because of slot blot...