Screening And Identification Of Differentialy Expressed Genes In Goose Hepatocyte Induced By Free Fatty Acid

Hepatic steatosis results from the excessive accumulation of triglyerides(TG) in the animal liver.FFAs cultured with Hepatocytes could induce Hepatoeytes steatosis, but it has been not clear about the relation between FFAs and TG abnormal deposition,and the mechanism of differentialy expressed genes in fat deposition.In this study,using the Sichuan White geese as material,we construct two forward and reverse SSH cDNA libraries between the normal hepatocytes and the steatosis hepatocytes.By homology alignment and function annotation,we discoverd the sorts and quantity of differentialy expressed genes in FFAs induced hepatocytes,which would provide a bias for elucidating the mechanism of fatty liver in the molecular level.The Results were shown as follows:1.The hepatocyte steatosis models were established successfully induced by 1mmol/ml FFAs for 36d.2.Differentially expressed cDNA library were constructed using suppression subtractive hybridization.1344 positive clones were obtained in SSH-1,1152 positive clones were obtained inSSH-2.SSH-1 library had 30 successful sequences,and SSH-2 library has 23 successful sequences.3.A search for sequence homology in the GenBank and EST database by BLASTn revealed that the distribution of SSH-1 EST clusters for high homology genes.The high homology genes,high homology ESTs and novel ESTs were23,6 and 1,and the proportion were 76.7%,20%and 3.3%.In SSH-2,the high homology genes and novel ESTs were 19,land 4,and the proportion were 78.3%,4.3%and 17.4%.4.the proportions of the high homology genes was 41 in SSH library.Categories of functions ESTs:the proportions of cell structure/movement was 2%,the cell signal/conduction was 12%,slation/expression regulation was 25%,transporters was 5%cell apoptosis was 2%, material conveying was17%,metabolismd and unknow gene were 22%and15%.Of course, the genes involved in metabolism and transporters were rich expressed.5.Roles of different character feature of partial diferentially expreesed genes in two geese breeds were presumed based on references and our research results.RP (RPL22,RPL23,RPL39,RPL5),TGM3,FABP,THBS1,and APOH were considered playing an important role in different character feature in different geese.They are the biological function of providing the material and they provide a clue in searching the important genes related to the hepatic steatosis and the foie gras.