Screening And Identification Of Differentialy Expressed Genes During Hepatocytes Of Landes Geese And Sichuan White Geese

Suppression Subtractive Hybridizition is a new and effective method to search differentialy expressed genes. In this study, the hepatocytes of Landes geese and Sichuan White geese were isolated as materials to search differentialy expressed genes between the two breeds. The information about differentialy expressed EST between the two breeds was got by screening and identification EST sequence making use of bioinformatics methods,The Results were shown as follows:(1) This study succeed to to segregate hepatocytes of Sichuan White geese and Landes geese using "Seglen two step collagenase dabbling". And by MTT detecting cell cytoactive, the result indicated, the cell cytoactive raised very slowly on the first to second day, And it begin to descend on the second to the third day.but the cell cytoactive raised very quickly on the third to the fifth day. Later it again began to depress. Change for cell cytoactive is same as the typical "S" growth curve.(2) In this differentially expressed cDNA library were first constructed using suppression subtractive hybridization between Landes geese and Sichuan White geese, and 2688 positive clones were obtained. The results showed that there were inserted fragments of 250-750 bp in the randomly selected positive clones, which indicated the sizes of inserted fragments for the libraries clones matched with expected consequence. The utility masculine rate for SSH-1 library and SSH-2 library reached 93% and 91%.(3) Differentially screening was performed to screen differentially expressed genes using DIG dot-blotting method. After part of clones were detected, sixty five strong signals clones were isolated and sequenced resulting in fifty four sequences.(4) A search for sequence homology in the GenBank nr and EST database by BLASTn revealed that the distribution of EST clusters for high homology genes, high homology ESTs and novel ESTs were 50, 2 and 2, respectively, in 54 ESTs. The high homology genes was grouped into 8 clusters based on the best known function of there protein products:signaling transduction/communication, cell structure/mobility, transporters, apoptosis, metabolism, translation/expression regulation, cell/body defence and one cluster that function is unknown or without an ontological theme. Of the total, the two type genes of specific gravity are very large which is participate signaling transduction/ communication and cell structure/mobility, and they were 16% and 28%, and the ratio of translation/expression regulation genes was 14%.(5)Roles of different character feature of partial diferentially expreesed genes in two geese breeds were presumed based on references and our research results. ACT, GST, CTGF, ELOVL, serine/threonine kinase, and elFI were considered playing an important role in different character feature in different geese, but its specific function, translation/expression regulation and signal pathway for the two geese breeds need to be researched furtherly.