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Host factors involved in Rous Sarcoma Virus replication
Štafl, Kryštof ; Svoboda, Jan (advisor) ; Horníková, Lenka (referee)
Rous sarcoma virus (RSV) takes a place of honor among retroviruses. Research of RSV allows us to uncover the secret of the origin and evolution of life, the mechanisms of tumorigenesis and the interaction between viruses and their hosts. Viruses are not able to replicate themselves without host cells. They exploit a number of cellular pathways and factors and they can reprogram the cells to produce great amounts of viral progeny. They exert pressure on their host that leads to developement of new types of cellular proteins, which then results in resistance of the cells. This thesis focuses on the host factors involved in the RSV replication cycle. It summarizes the current knowledge about the replication cycle of retroviruses and the host factors that are necessary for productive infection: cellular receptors, endocytic and secretory pathways, nuclear transport, proteosynthesis, replication of proviruses and its stimulation. The restriction mechanisms of cells are also taken into account. The current knowledge about RSV is compared with facts on mammalian retroviruses and gaps in research are highlighted. The influence of the host cell factor absence, host specificity and cellular permissiveness are correlated and discussed.
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Detection of phytoplasmas using DNA-microarrays
MARKOVÁ, Jaroslava
The aim of this thesis was to optimize the method of detection of phytoplasmas using DNA-microarray. It consisted of testing an appropriate method of genetic material isolation, development and optimization of PCR to amplify different groups of phytoplasmas, optimization of detection of DNA at a microarray, and sequence analysis of phytoplasma in order to design more suitable probes. PCR was first optimized for collection isolates, then also for natural samples. All 16Sr groups from the collection were sequenced and phytoplasmas were detected in them by hybridization. Phytoplasmas were detected also in natural samples: oilseed rape (species Brassica napus), red clover (Trifolium pretense), purple coneflower (Echinacea purpurea), and apple tree (Malus domestica). Using the DNA from insect vectors, only sample 202/6 from the group 16Sr-XII was positive. The sequence of red clover and oilseed rape correspond with the database samples in the group 16Sr-I "Aster yellows".
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