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Proteomic differentiation of hairs of different dog breeds
Humpoláková, Karin ; Kučková, Štěpánka (advisor) ; Kolář, Karel (referee)
The goal of my thesis was to find out whether it is possible to distinguish among individual dog breeds from the protein analysis of their fur. This knowledge could be used, for example, in forensic science. In this work, there was also a comparison of the similarity of dog breeds with a wolf, which was domesticated and is considered as an ancestor of the dog. For this study, the hair of three representatives of sixteen dog breeds was collected. To analyse these samples enzyme cleavage was used a trypsin, and mass spectra were obtained by MALDI- TOF MS (Matrix-Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry) and LC/MS-MS (Liquid Chromatography with Tandem Mass Spectrometry) methods. The obtained data were evaluated by the PCA (Principal Component Analysis) and LDA (Linear Discriminant Analysis) method. It was found, that individual dog breeds cannot be distinguished using both methods. KEYWORDS proteomics, coat, dog, breeds, mass spectrometry
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Identification of animal hair by mass spectrometry
Humpoláková, Karin ; Kučková, Štěpánka (advisor) ; Kolář, Karel (referee)
The goal of this bachelor work was to find out if it is possible to recognize mammal species by analyzing the proteins extracted from their hairs. This information could be of use, for example, in forensic science as evidence in cases of exotic animal species smuggling or it could be helpful for solving some specific criminal cases. Fur is a characteristic sign of mammals which is forming their body surface. It mainly consists of keratin, which is a fibrous protein. This bachelor thesis deals with proteomic analysis of animal hair using mass spectrometry working on MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization - Time of Flight) principle. Analyzed animal hairs were collected from ten local domestic and wild animals, such as dog, goat, cat, fallow deer, sika deer, degu, horse, rabbit, roe deer and Belgian blue calf breed. For the analyses of samples that are rich in proteins a specific enzyme cleavage by trypsin was used. The proteins were cleaved into peptides, from which mass spectra were obtained by MALDI-TOF mass spectrometry. Then these spectra were evaluated by the Principal Component Analysis (PCA) method. Then the obtained aminoacid sequences of the keratins found in the individual animal hair were searched through publicly available protein database - UniProt. The aminoacid...
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