Abstract (eng)
Understanding of the regulation of the viral life cycle is important to develop tools for genetic manipulation or protein synthesis in the host organism. This is especially important for less studied organisms. This thesis therefore concerns the transcriptional regulation of the virus φCh1, which infects the haloalkaliphilic archaeon Natrialba magadii.
The first part of this thesis deals with the regulation of the tail fibre protein gp34. This protein is a part of a phase variation system, in which the C-terminal end can be exchanged to produce two different orientation of this protein. Production of each variant in the cured strain Nab. magadii L13, showed that the (+) orientation reduces the infectivity of φCh1. This result is another confirmation that only one orientation of gp34 is able to adsorb to Nab. magadii. It has been further shown, that the lysogeny regulators ORF48 and ORF49 have no influence on the production of the tail fibre protein. However, ORF44 has a repressing effect on ORF34. ORF44 is a part of a putative archeal toxin- antitoxin system and has homologies to a PIN-domain nuclease. The antitoxin of ORF43 prevents this repressing effect.
The focus of the second part of this thesis lies on the ORF34 regulator ORF79. Previous experiments revealed a repressing activity of ORF79 on the production of gp34. Here, the specificity of the regulator was tested by co-expression with other φCh1 genes. The first investigated gene was soj. In the course of this experiment an antibody against the protein Soj was created. It has been shown that the production of this protein is not regulated by ORF79, but this protein had an influence on plasmid stability. This could be explained by the homologies of Soj to partitioning proteins. For this reason the influence of ORF79 on a second protein was observed. Therefore, the methyltransferase was used, which is also not affected by ORF79. Additionally, the expression pattern of ORF79 was monitored and compared with the expression pattern of ORF34.
The last part of this thesis deals with the regulator ORF49, which is involved in the switch between lysogenic and lytic life style. To further characterize this ORF, I tried to construct a homozygous deletion mutant. This attempt failed, because the complete deletion of the wild type ORF49 was not possible.