Application of the SDS-PAGE method for the characterisation of winter wheat flour quality112-118Views:98
The 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.
Qualitative detection of genetically modified organisms in plant samples309-313Views:90
We analysed the GMO content of corn samples by polymerase chain reaction following the appropriate optimization of the reaction. The analysis included two main steps: extraction of DNA from the sample, and detection of the GMO content by polymerase chain reaction. The polymerase chain reaction is an in vitro method to multiply chromosomatic or cloned DNA (cDNA) sequences through the enzymatic pathway. The reaction is sensitive enough to produce DNA in sufficient amount for the analysis from a single DNA. We identified the PCR products by agarose gel electrophoresis. When optimizing the reaction, the MgCl2 concentration, reaction time and temperature have to be taken into consideration. The temperature of the anellation has to be increased until the highest specificity and yield is reached. If the temperature of the anellation is too high, the primer is linked to non-specific sites as well; in the gel visualization, more lines can be seen at one sample. If the temperature of the anellation is too high, the primer is insufficiently linked or is not linked at all (too few lines in the gel visualization). After optimization, the GMO content in the unknown sample can be determined along with the appropriate positive and negative controls.
The composition of gluten proteins and their effect on the rheological properties of gluten124-129Views:117
Wheat is the major cereal component of bread in the world and is grown worldwide. Of the cereals only the bread wheats – and less the triticale – includes storage proteins that play an important role in the performance of gluten. Proteins of gluten complex may be present in two classes:
− low molecular weight (gliadin-) components, and
− high molecular weight (glutenin-) components.
Gliadins shown appreciable heterogenity and can be separated into 40-50 components with gel electrophoresis. The composition of gliadins is employable for the identification the wheat varieties and to investigate the varieties. In the decreasing electrophoretic mobility sequence may be distinguish α-, β-, γ- and ω-gliadins. A glutenin subunits may be include in two classes:
− high molecular weight glutenin subunits (HMW-GS),
− low molecular weight glutenin subunits (LMW-GS).
Wheat varieties can be identified by glutenin and their quality selection is also possible. The gliadin’s polypeptides encoding genes are located on the short arm of chromosomes 1A, 1B and 1D, 6A, 6B and 6D. Genetic coding for HMW subunits is located on the long arms of chromosomes 1A, 1B and 1D, the LMW-GS are also located on chromosomes 1A, 1B and 1D (Glu-3 loci) near the gliadin-coding loci.
Storage proteins affect the rheological properties of gluten by two factors:
1. The quality and quantity of the protein components of the gluten complex,
2. The interactions between the protein fractions.