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Application of the SDS-PAGE method for the characterisation of winter wheat flour quality
112-118Views:547The principle, development and importance of the SDS-PAGE method are presented in this article. The SDS-PAGE method has become one of the basic methods of molecular biological research, because it is widely applicable and its sensitivity is excellent in the separation of wheat storage proteins.
We have shown the application of this method with a concrete example. It was also tested whether, it was possible to obtain a better baking quality product from a large amount of poor quality less valuable wheat by fractioning the flour according to particle sizes after grinding. We studied the rheological properties of flours with different particle size fractions from the original flour. The baking quality of the original flour was B2. The 125-90 and 90-63 μm fractions have significantly better baking quality (B1) than the original flour. The protein contents of these flour fractions were also significantly higher than the protein content of the original flour. We had a question: what has influenced the baking quality: the protein content or other factors? We searched for an explanation on these results in the protein composition of the flour samples. We studied the distribution of glutenin-fractions by SDS-PAGE method and evaluated them. We found with correlation analysis that the amount of LMW-Glutenin D-group (52-60 kDa) is in a strong, negative correlation to the baking quality (r = – 0.855*). Therefore, the baking quality of flour samples was influenced by this glutenin fraction. -
Analysis of longevity in Holstein Friesian cattle using proteomic approaches
21-25Views:639The aim of the present study was to determine marker proteins those are associated with functional longevity of dairy cattle. Holstein-Friesian cows were grouped based on their performance as follows: group 1) individuals with good longevity traits; group 2) short production life because of poor reproduction traits; group 3) short production life with low milk yield. Twelve individuals were sampled in each group, blood and milk samples were collected from cows. Blood samples were analysed with two dimensional polyacrylamide gel electrophoresis (2D PAGE), MALDI TOF/TOF and nanoLCMS/MS. The milk samples were analysed with MALDI TOF/TOF and nanoLC-MS/MS. Using the optimized gel based proteomic approach,
we have succesfully separated 143 proteins in the group1, 139 proteins in the group2 and 136 proteins in the group3, but we could not find significant differences between groups in the expression pattern. Using MALDI TOF/TOF and nanoLC-IonTrap MS, we have found eleven protein sequences those were expressed only in the samples of good longevity group. -
Effect of selenium supplement on proteome of chicken liver
9-13Views:561The aims of the present study were to optimize a two dimensional polyacrylamide gel electrophoresis method to chicken liver proteome and to determine the changes of protein expression caused by selenium. Twelve broiler chicken were used in this experiment. The selenium intake was 0,2 mg/kg in the control group (6 chicken) and 4,25 mg/kg in the experimental group (6 chicken). Using the optimized proteomic approach, we have succesfully separated 747 proteins in the experimental group and 741 proteins in the control group. We found six proteins, wich expressed only in the samples of experimental group. Further investigations need to determine these six proteins with mass spectrometry (MS) and look for the correlation between the physiological effects of selenium and the expression of these proteins.