Carbohydrate-related Gene Chip Design And Preparation

Background:Glycomics is one most advancing sphere in life science.Like nucleic acid,the saccharide also perform a very important part in the extension of gene message,so the research in glycomics apears extremly urged during the developing of genomic and protemic research.The oligosaccharide chains in glycoproteins and glycolipids often apperance in the cell surfaces and the proteins which secreted external cell.The oligosaccharide chains paly an impotant way not noly in effecting the functions of glycoprotein,but also regulating with cell-recognition,signal transmission,endocytosis,cell growth,cell differentiation and apoptosis.The construction and amount of oligosaccharide chains are changing during some diseases happening.The synthesis of oligosaccharide chains is catalysised by gene-codogenic glycotransferases,it is considered that about 0.1～1.0%genes in mammalian genome participate the synthesis and metabolism of oligosaccharide chains.The reaserch of genetic expression of a whole set of enzymatic system which take part in forming N- and O-oligosaccharide chains is very helpful in revealing the relationship between the glycogene and glycostain,it is also significance in a diagnostic way.Purpose:Gene chip is a high-flux detecting method emerging decades years.our essay adopt an assay preparative method by 60-mer oligonucleotide.we shift the human glycosyltransferase,glycosidases and sulfotransferases from the database of CFG(Consortium for Functional Glycomics) and CAZY(Carbohydrate-Active enZymes ),then pick up the mRNA sequences in the GeneBank.By using the Oligo 6.0 and BLAST,we can design probes for the hybridization properly.Further more,after printed on the amino glass when the 60-mer oligonucleotide probes are ready,we can get the finished products by others steps.Conclusions:We have designed hundreds of 60-mer oligonucleotide probes,which compose 130 glycosyltransferase genes,12 glycosidase genes,48 sulfotransferase genes and 10 housekeeping genes.we prepare the glycogene chip and certificate its reliability by more experments;Besides,we take a comparative study in the KBD blood and normal blood by home-made gene chip,and find out 49 abnormal glycogene which comprise 28 up-regulative genes and 21 down-regulative genes.We picked up 2 genes those expressing discrepancy deeply and taked relative quantitions by Real-Time PCR.The result shows that it conforms to the result from the glycogene chip.This research provides a novel glycosylgene chip as a technical paltform to indicate the relationship between the diseases and glycogenes.