Refine
Document Type
- Master's Thesis (4)
- Bachelor Thesis (1)
Keywords
- Sequenzanalyse <Chemie> (5) (remove)
After the expression of the titin-Hsp27-construct with the following purification supplies no satisfied results which makes the realization of the atomic force microscopy not possible. The devel-opment of the structure model by using different bioinformatic methods can establish a model for the protein sequence. As bioinformatic methods the template search by different BLAST runs and free available software like SwissModel, Pcons, ModWeb and other tools are used. Nevertheless, the generated model is not the native conformation and has to be analyzed with other software until a stable conformation of the structure can be predicted. Depending on the time which is provided the generated model is a good approach for the aim this master thesis has.
Die vorliegende Arbeit befasst sich mit der Sequenzanalyse der für die Legionellose relevanten Schlüsselproteine der Organismen Legionella anisa, Legionella drancourtii und Legionella shakespearei, sowie mit in vitro-Kultivierungsexperimenten der Spezies Legionella pneumophila ATCC 33152. Zunächst wurden die gesetzlichen Gegebenheiten der Legionellenproblematik und die biologischen Hintergründe betrachtet, darunter die intrazellulären Mechanismen der Legionellose. Die Ergebnisse der Arbeit beinhalten Informationen über die Schlüsselproteine der drei Legionellen Spezies und deren mögliche Pathogenität.
In the present bachelor thesis, nanopore sequencing and Illumina sequencing was compared using pollen DNA collected from honeybees and bumble bees. Therefore, nanopore sequencing was performed with the MinION sequencers and the generated reads were analysed with bash programming. A quantitative and qualitative (based on ITS2 sequences) BLAST run was performed. The results confirme the error probability of nanopore sequencing that is described in the literature. Nevertheless, with both sequencing methods similar sample preferences of the bees could have been observed, allowing ecological conclusions.
In this work, a protocol for portable nanopore sequencing of DNA from pollen collected from honey bees, bumble bees, and wild bees was developed. DNA metabarcoding is applied to identify genera within the mixed DNA samples. The DNA extraction and ITS and ITS2 PCR parameters tested for this purpose were applied to the collected pollen sample and the amplicons were then decoded using the Flongle sequencer adapter from Oxford Nanopore Technologies. It is shown that the main pollinator resources at the different sites can be identified in percentage proportions. The protocol generated in this study can be used for further ecological questions.
RNA tertiary contact interactions between RNA tetraloops and their receptors stabilize the folding of ribosomal RNA and support the maturation of the ribosome. Here we use FRET assisted structure prediction to develop structural models of two ribosomal tertiary contacts, one consisting of a kissing loop and a GAAA tetraloop and one consisting of the tetraloop receptor (TLR) and a GAAA tetraloop. We build bound and unbound states of the ribosomal contacts de novo, label the RNA in silico and compute FRET histograms based on MD simulations and accessible contact volume (ACV) calculations. The predicted mean FRET efficiency from molecular dynamics (MD) simulations and ACV determination show agreement for the KL-TLGAAA construct. The KL construct revealed too high FRET efficiency and artificial dye behavior, which requires further investigation of the model. In the case of the TLR, the importance of the correct dye and construct parameters in the modeling was shown, which also leads to a renewed modeling. This hybrid approach of experiment and simulation will promote the elucidation of dynamic RNA tertiary contacts and accelerate the discovery of novel RNA interactions as potential future drug targets.