Establishment Of 2-DE Map And Identification Of Differential Protein Of Bovine Whey From Healthy Cows And From Those With Clinical Mastitis By LC-MS/MS

Whey protein expression pattens of healthy cows and those with clinic mastitis were investigated by using two-dimensional gwl electrophoresis (2-DE) and LC-MS/MS in the current research to study the mastitis-associated protein changes.. The main results obtained are presented.1. The optimal measurement condition for separating proteins of whey from clinical mastitis and healty dairy cows was established by using the two-dimensional gel electrophoresis (2-DE) colloidal stained with Coomassie and silver, and the normal bovine milk whey proteins and mastitic bovine milk whey proteins were separated successfully.2. 142±4, 82±3 and 111±6, 80±4 normal whey proteins and mastitic bovine milk whey proteins were separated into the the areas of pl 3-10 using Coomassie stained gel and silver stained gel, repectively. Therefore, better resolution and repetitive 2-DE atlas were obtained, the optimized 2-DE and image analysis technique is a better method to support further study of whey from cows infected with clinical mastitis and healthy dairy cows.When coupled with 2-DE, silver stained gel and Coomassie stained gel, there were 31 and 13 proteins expression of normal whey and mastitis whey showed clear differences, respectively. Furthermore, the patterns of proteins expression was repeated 3 times, highlights the changes which take place during clinical mastitis with the number of 7 and 6 proteins expression increased, but the number of 24 and proteins expression 7 observed decreased by analyzing the image of 2-DE silver stained gel and Coomassie stained gel, respectively.3. The identifications of the numbered protein spots by MALDI-MS which whey obtained from clinical mastitis and healthy dairy cows, getting the four differences proteins: casocidin-I; Chain F, Crystal Structure Of Bovine Alpha-Lactalbumin; Chain B, Bovine Beta-Lactoglobulin, Lattice; Structure Of Diferric Bovine Lactoferrin At 2.8 Angstroms Resolution being identified. Probably these 4 proteins have a key influence on the mechanism of mastitis and provide a new understadingc for the treatmet clinical mastitis by using a biological approaches.