| Organic arsenic preparations, such as arsanilic acid, used widely to promote the development of animal husbandry, and achieved good social and economic benefits. However, animals are apt to arsenic accumulation, long-term feeding arsanilic acid, lead arsenic residues in animal products exceeded. The problem of veterinary drug residues in meat has attracted wide attention. Nowadays, researchs are basically focused on the rapid screening analysis and validation of analytical detection methods, but these studies did not elaborate on the fundamental veterinary drug residues in meat, poultry and then changes in the body caused by a variety of biological mechanisms. Therefore, an urgent need to find a method of analysis, the system can be intuitively reflect the body of veterinary drugs in animal metabolism. Proteome research for pork liver traits will allow scientists to build and test better hypotheses. Two dimensional gel electrophoresis (2-DE), treated as the important proteomics technique, enables the separation of complex mixtures of liver proteins according to isoelectric point, molecular mass, solubility, and relative abundance. In this study, protein expression of piglet liver analysis to explore the organic arsenic preparations (arsanilic acid) in pigs biological mechanisms by the use of two-dimensional electrophoresis, which essentially describes the kinds of veterinary drug residues in meat for the impact. To be able to more directly reflect the preparation of organic arsenic metabolism in the animal body to properly control the process of veterinary drugs in pig breeding applications, provides a new way of thinking.1. Establish proteome analysis of pork liver by two dimensional gel electrophoresis. Obtain high quality and reproducible two dimensional gel electrophoresis maps. Based on the pork muscle protein characters, the related program of2-DE was established as follows:sample preparation by ultracentrifugation and Clean up kits,17cm pH5-8IPG strips, and protein sample volume120μL. The results showed that the matching rate was76%±1.7%and about1412different protein spots were observed.2. Blind method was used to verify the accuracy and repeatability of the above system. The matching rates of different pork livers were more than70%, and the midpoint of the relative migration rates were within the laboratory control standards. As for the protein detection, the method optimized in the present study was found in both significant statistics and significant biology. 3. Single-factor completely randomized experiments under the arsanilic acid on liver protein of piglets. The maps between the control group and test group piglets were with good reproducibility and stability. Compared with PDQuest software for variance analysis, the results obtained, both statistically significant and analysis significance of differences in protein spots13, all spots were raised proteins.4. The13differentially expressed proteins points of the control group and test group piglets were sent to MS+MS/MS analysis, using MASCOT software in the protein database to search matched protein,12points were successful identified. Obtained10kinds of proteins, respectively, were Chain B, Refined1.8Angstroms Resolution Crystal Structure Of Porcine Epsilon-Trypsin, Rho GDP dissociation inhibitor alpha, PREDICTED:hypothetical protein LOC100512997, PREDICTED.-26S proteasome non-ATPase regulatory subunit13-like, PREDICTED:LOW QUALITY PROTEIN:heat shock cognate71kDa protein-like, PREDICTED:synaptic vesicle membrane protein VAT-1, cytoskeletal beta actin, PREDICTED: actin,cytoplasmic1, PREDICTED:actin,cytoplasmic, cathepsin B precursor. Analyze the function of these proteins to explore the arsanilic acid on piglet liver protein expression, found that are closely related a series of the metabolism in the liver and damage to the body by arsanilic acid. |
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