Screening And Primarily Characterizing ESTs Differential Displayed In New-born Black-Bone Chicken's Liver

Black-bone chicken is one kind of unique domestic chicken breeds and widely spreads in China. Owing to the Silkies' unique traits, high officinal, viewing and nutritious values, scientists put high premium on its breeding and exploitation for a long time. The essences of variations among breeds' traits or individuals' traits are subject to the different kind of expressing genes or the same gene but in differently expressing amount. Research on discrepancy in gene or gene's expressing amount is a useful way to obtain the information of life and uncover traits' occurrence and development. Furthermore it can also provide important information of which how the unique traits of black-bone come into being and form the base of advanced research of black-bone chicken.Excluding interference of breeding conditions' distinctness, this trial compared the difference of gene' expressing in liver which closely correlated with Silkie's, normal plumed black-bone chicken's, SuQin96's and Abar acer broiler's unique traits.By using differential display reverse transcription PCR that piloted by combinations of three anchor primers and twenty-four random primers, we obtain 251 different expressed fragments, which were cut from polyacrylamide gel electrophoresis products of four breeds'cDNA. With Re-PCR, electrophoresis and purification, we got 220 different expressed fragments. After SSCP detecting, there were still 198 singularities. The singularity rate is 90%. Through gene chip scanning and comparing comprehensive data of signal value, we confirmed eight fragments were genuine differential display fragments. We only attain 4 sequences by cloning and sequencing at last. The reason might be the differential display fragments' low amplification rate. And what's more, they were short in length and feeble in brightness. We thought the reason lays in the differential display genes' low abundance.The four ESTs were cleared contaminated vector sequences up and submitted to NCBI.Now they were granted accession number: DQ003271, DQ003272, DQ003273, DQ003274.These ESTs were chromosomal assigned by BLAST with chicken genome: DQ003271, located at chromos 5; DQ003272, located at chromos 3; DQ003273, located at mitochondrion; DQ003274, located at chromos 19. DQ003271 was a chicken's new EST whose function was supposed to be similar to coiled-coil like protein 1 .Three others were existent chicken ESTs. DQ003272 was induced to influence organism immunity.DQ003273 may be a vital factor that was related with cell redox. And DQ003274's function was uncertainty.Liver is 'Metabolization Centrum'. What it expressed accounts for overwhelming percentage of what organism genes expressed in a certain time and a certain tissue. The diversity of these ESTs' functions reflects the diversity of liver 'function and it's importance in whole organism.These results of trial provide significant data for further research on differential traits among black-bone chicken, layer and the broiler. However there needs in-depth experiment design and analysis on each EST and the gene it represents fixing on their structures, functions and relations between traits and differential display genes.