| Diabetes is a disease resulting from relative or absolute insulin deficiency. For both type I and type II diabetes, the major clinical problems result from the long-term effects of chronic hyperglycemia. Recent results from the Diabetes Control and Complications Trial (DCCT) demonstrate that tight glucose control lowers the incidence of diabetic complications, which are the major causes of morbidity and mortality from diabetes. The ultimate destination for diabetes mellitus treatment will be one that maintains a normal blood glucose concentration in the face of fluctuating dietary intake. Unfortunately, intensive diabetes management involving many injections per day results in an increase in episodes of hypoglycemia, so it is not a ideal treatment. The promising strategy for diabetes management is gene therapy, which can be broadly defined as genetic modification of cells by using gene transfer technology to recover insulin physiological release and to improve the clinical status of patients with diabetes. Genetic engineering of non-β -cell to substitutes sick β -cell of patient is a strategy of gene therapy for diabetes. A potential advantage of using cells other than β-cell is that non-β-cell might not be recognized by the autoimmune response directed against β -cell in patients with type I diabetes. The difficulty of this approach is to find an ideal gene promoter. To study the effect of phosphoenolpyruvate carboxykinase(PEPCK) gene promoter on regulation of reporter gene transcription, we conducted the following works:Firstly, Construction and identify of recombinant firefly luciferase reporter plasmid pGL2-PEPCK-Luc. A 550bp fragment of PEPCK promoter cut from plasmid pPEPCK-int was inserted into transitional vector PBS-SK to construct a transition plasmid PBS-PEPCK. Then, Basing on this plasmid we successfully cloned the recombinant luciferase reporter plasmid pGL2-PEPCK-Luc. By surveying the nucleotide sequence conformed that the...
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