Application Of2-DE Technique For Differences In Chick Serum Protein Analysis Before And After Cold Stress

Stress factors are various,In the north,especially in the winter,cold stress is animals（especially chicks） of the most common stress factors,seriously affecting livestock andpoultry production performance and disease resistance. Serum have rich protein,they may bereacted physiological state of normal physiological state and disease situations, it may beimportant to the diagnosis of a disease marker protein,as is further also possible drug targets.Therefore,it is of great significance that looking for high specificity,high sensitivity to coldstress markers by the animal body fluids such as blood,tissue,etc. The use of high-throughputproteomics analysis and detection of protein expression differences in serum levels of chickenunder cold stress conditions,could get a change of key proteins in the influence of cold stress，helps to reveal and validate protein with specific physiological, pathological processes relatedto cold stress and explore the molecular mechanism of the development occurs.On the basis of this study to establish the serum proteome removal of high-abundanceproteins,thereby reducing the high-abundance proteins of low abundance proteins shelter andincrease the sensitivity of separation technology. Select cold stress and cold-adapted group ofchicks serum as the main object to room temperature as a control group of chicks serum,andcombines two-dimensional gel electrophoresis（two-dimensional gel electrophoresis,2-DE）;using image analysis software PDQuest8.0analyzed cold stress serum proteomics andcold-adapted group and normothermia group differences in expression profiles images. Afterthough mass spectrometry identification of cold stress-related proteins.First,preliminary analysis serum of30chicks,using two-dimensional gel electrophoresisseparation and Coomassie blue staining to examine the quality of sample preparation,optimization and comparison of two-dimensional electrophoresis conditions and the removal ofhigh-abundance proteins. Successfully established chick serum protein two-dimensional gelelectrophoresis. After chicken serum preliminary gap analysis,Matrix-assisted laser desorptionionization time of flight mass spectrometry （MALDI-TOF-MS） to identify the differenceproteins and Western blot verification.The results showed that: Through2-DE analysis normal temperature,cold stress andcold-adapted treatment serum chicks,obtain a relatively complete differential proteins data and found23differentially expressed proteins. After the analysis found that protein expression levelexpression increase and expression down points of difference were11and12.MALDI-TOF-MS analysis8distincts and reproducible differences protein spots,finalsuccessfully identified4different proteins. Obtained different proteins are related proteins ofglucose and the energy metabolic pathways,ALDOC protein plays a crucial and pivotal role inthe glycolysis and fructose metabolism, it may also occur in cold stress and resistance to coldstress for the body for energy play an important role. Subsequently,the Western blot method fordifferential protein ALDOC further validation, the results were consistent with the results of the2-DE.