|
|
Optimalization of PCR-RFLP method for taxonomy of yeasts
Olivová, Radana ; Vadkertiová, Renata (referee) ; Vránová, Dana (advisor)
This thesis deal with optimalization method PCR – RFLP for taxonomy enlistment of yeasts. Conventional identification methods for yeasts are time-consuming. Molecular biological method based on PCR are instrumental towards fast and precise identification as compared to conventional phenotypic methods. In this thesis molecular biological method PCR – RFLP was used for identification and enlistment of yeasts. This metod follow repeating spacers of ribozomal DNA of yeast, characteristic for each species and strain. By the help of PCR were amplified specific partitions of DNA. These fragments of DNA were split by restriction endonucleases and identified by horizontal electroforesis. In background of this thesis there are information about yeasts, their taxonomy and molecular biological methods.
|
|
|
Identification of wine yeasts by PCR-RFLP method
Šuranská, Hana ; Vojtíšková, Marie (referee) ; Vránová, Dana (advisor)
This thesis deals with identification of the wine yeasts by applying the PCR-RFLP method. The identification and characteristic of the yeasts has gone through substantial changes in recent years. There have been introduced new methods of taxonomic classifying based on the molecular methods, which are oriented to easy and fast identification. One of these methods is the PCR-RFLP method. The amplification of the 5•8S-ITS rDNA sequence by the polymerase chain reaction with use of the primers ITS1 and ITS4 leads to the amplification of the specific sequence of DNA. Such multiplied DNA is after repurifying by the ethanol and drying submitted to the restriction analysis. With use of the restriction endonuklases DNA is chopped into the specific segments typical for the particular genus. The chopped fragments can be separated in the electric field in the agarose gel and subsequently evaluated. In this thesis together 63 type yeasts were used. These yeasts were analysed by applying of the seven restriction endonuklases – HaeIII, HhaI, HinfI, HpaII, TaqI, AluI a MseI. The final image of type yeasts splitting was compared to the results of splitting of already identified wine yeasts and these yeasts were subsequently taxonomically classified. Evaluation of genetic similarity was conducted by program BioNumerics and as the results the dendrograms that were created with use of Jaccard‘s coefficients are obtained.
|
|
|
The identification of Saccharomyces yeasts during fermentation of white grape must.
Zdeňková, Michaela ; Vojtíšková, Marie (referee) ; Vránová, Dana (advisor)
This thesis is concerned with the identification of the yeasts belonging to Saccharomyces species, which is participating in the particular fermentation phases of the white wine. The rapidity that we are able to identify the yeast strains is the significant factor for appreciation the fermantation proces quality as well as final product quality – the quality of wine. The molecular biology method developing is gradually limiting the using of traditional identification methods mainly because of their time intensity. In this thesis was used molecular method PCR-RFLP as an implement to quick and accurate identification of particular strains of yeasts. The specific DNA section was amplified by the help of PCR and consequently amenabled to the restrict analysis. The restrict endonukleas fissiled DNA to fragments specified for the certain strain of yeast. The fragments were detected by horizontal electrophoresis. To compare the fragments with type yeast fragments made us possible to identify the trast strain and its taxonomy classification. The literature search contains the basic information about the yeasts and their using, the information about wine making as well as the PCR-RFLP method principle.
|
|
|
Identification of yeasts genus Saccharomyces from leaves, grapes and must of grape-vine.
Škodová, Anna ; Štros, Michal (referee) ; Vránová, Dana (advisor)
This diploma thesis is concerned about the PCR-RFLP method for identification of yeasts genus Saccharomyces from leaves, grapes and must of white wine. The thesis describes an obtaining of pure cultures of yeasts, isolation DNA and amplification of specific segments of DNA using PCR method. Splitting of this segments by restriction endonukleases and a detection of final fragments by zone electrophoresis are mentioned too. Length of fragments and numbers of restriction fragments, which are characteristic of every species, enable a determination of microorganisms and their inclusion into the system. The basic informations on yeasts and molecular methods for their identification are named in the theoretical part of the thesis. The main part of the thesis deals with the processing of grapes of grape-wine and the manufacturing of white wine.
|
|
|
Taxonomic submission of Saccharomyces yeast from selected matrices
Mašitová, Lucie ; Vadkertiová, Renata (referee) ; Vránová, Dana (advisor)
This thesis explores the optimizing of the methods relevant to the cultivation, isolation, and identification of individual yeast strains from selected matrices, using the molecular biology methods. Grape berries, vine-leaves and soil from vineyard have been used as matrices. As yeasts are an integral part of fermentation processes, they are used for making of wine, the organoleptic charakteristics of which they influence. For identification of yeast strains the PCR-RFLP method has been used. Specifity of yeast DNA sequences of certain species has been used for this analysis. These spacers have been amplificated through the use of PCR and consequently they have been subjected to a restrictive analysis with specific restrictive endonucleases. These fragments have been detectioned through horizontal electrophoresis. In the literature background research section the basic informations about viticulture, yeasts, their cultivation and separation, PCR, restrictive analysis and horizontal electrophoresis is presented.
|
|
| |
|
| |
|
|
Study of historical expansion of brown trout lineages in the Czech republic and in Slovakia republic with using mtDNA markers.
JAŠKOVÁ, Iva
The brown trout (Salmo trutta) is an ecologically, economically, aesthetically fish species whose poor conservation status in the European countries calls for further attention and action. The continuing erosion of the genetic resources of brown trout populations by human activities calls for strategies to reverse the current trend. We studied a genetic diversity of population of brown trout in the territory of Czech R. and in Slovakia using genetic markers. In the fragments of mitochondrial DNA (gen for ND-5/6) and nuclear DNA (gen for LDH1) amplified through PCR the differences were searched with the use of RFLP. In tested populations seven haplotypes were founded, four haplotypes were represented in almost of all populations. The ``Danubian haplotypes{\crqq} were strictly confined to the Danubian and Vistula drainages, the ``Atlantic haplotypes{\crqq} dominated in all populations, most of the total molecular variance (72 %) was attributed to differences within populations. Two alleles at LDHC1٭ - the ancestral ٭100 and ٭90 (at a high frequency) were revealed. This genotypic replacement is considered to be due to anthropogenic activities.
|
|
|
Utilization of PCR-RFLP approach for evaluation of ITS polymorphism in selected \kur{Beauveria bassiana} strains/isolates
KOCKOVÁ, Lucie
The aim of this study was the evaluation of the suitability PCR-RFLP technique and ITS region analysis for determination the genetic variability of the entomopathogenic fungi - Beauveria bassiana, Beauveria brongniartii and Isaria fumosorosea. By the means of ITS-RFLP technique, it was possible to identify the uniformity of the B. bassiana isolates from NP Šumava, the higher level of polymorphism of B.bassiana isolates from different geographical areas was detected and it was possible to distinguish different entomopathogenic fungi used in this study.
|
|
|
The analyse of polymorphisms of chicken
TYLLEROVÁ, Helena
Chicken growth hormone gene was examined in chosen strains from DOMINANT CZ breeding program - Rhode Island Red (RIR), New Hampshire (NH) and the final hybrids RIRxNH and NHxRIR. Polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) analyses reveal three MspI restriction sites of intron 1 region. These restriction sites give rise to six RFLP fragments {--} 540 bp, 390 bp, 270 bp, 240 bp, 150 bp, 120 bp. Based on restriction patterns, three alleles of growth hormone were determined. From a possible six genotypes, only four were found in all populations. The allele A1 is the most abundant in RIR strain (0.925), allele A3 is the most abundant in NH strain (0.550). The allele A1 was present in the most of the hybrids.
|
guest ::
