Preliminary Analysis On Ratinal Gene Expression Profile Of Diabetic Rat

Objective 1. Investigating the feasibility of restriction fragment differential display—Polymerase Chained Reaction(RFDD-PCR) for a scan of gene expression in diabetic retina, We carry out this technique to establish the gene expression profile of nondiabetic and diabetic rat, to analyze and compare differences between them. 2 Investigating the expression of candidate genes selected by RFDD-PCR in different stages, to discuss their possible functions.Methods 1 The gene expression profiles of nondiabetic and 8-week Alloxan induced diabetic rat's retina were constructed by RFDD-PCR technique at equal pace according to operating manual of Qbio-gene company. 2. Then all fragments of the three gene expression profiles havebeen separated and displayed by electrophoresis on 7% urea denaturing polyacrylamide gel and scanned with Typhoon9200 Image system. Image analysis of the gels were carried out by using the software of ImageQuant TL, Image Tool, Fragment Analysis. 3. Bioinformatics analysis of retinal gene expression were performed by using the data means offered by gene database at the website http://www.Qbiogene.com/displav. Through comparing both expression profiles, the related gene fragment of diabetic retinopathy were selected initially as the target gene of further approach. 4 Manipulation was duplicated by selecting "expression windows" randomly. 5. The expression levels of selected candidate genes were identified by semi-quantitative RT-PCR in 4-week, 8-week and 24-week diabetic rats.