The genomes of diverse organisms are predicted to contain 20 – 30% membrane protein encoding genes and more than half of all therapeutics target membrane proteins. However, only 2% of crystal structures deposited in the protein data bank represent integral membrane proteins. This reflects the difficulties in studying them using standard biochemical and crystallographic methods. The first problem frequently encountered when investigating membrane proteins is their low natural abundance, which is insufficient for biochemical and structural studies. The aim of my thesis was to provide a simple method to improve the production of recombinant proteins. One of the most commonly used methods to increase protein yields is codon optimization of the entire coding sequence. However, our data show that subtle synonymous codon substitutions in the 5’ region can be more efficient. This is consistent with the view that protein yields under normal conditions are more dependent on translation initiation than elongation. mRNA secondary structures around the 5’ region are in large part responsible for this effect although rare codons, as well as other factors, also contribute. We developed a PCR based method to optimize the 5’ region for increased protein production in Escherichia coli. For those proteins produced in sufficient quantities several additional hurdles remain before high quality crystals can be obtained. A second aim of my thesis work was to provide a simple method for topology mapping membrane proteins. A topology map provides information about the orientation of transmembrane regions and the location of protein domains in relation to the membrane, which can give information on structure-function relationships. To this end we explored the split-GFP system in which GFP is split between the 10th and 11th β-strands. This results in one large and one small fragment, both of which are non-fluorescent but can re-anneal and regain fluorescence if localized to the same cellular compartment. Fusing the 11th β-strand to the termini of a protein of interest and expressing it, followed by expression of the detector fragment in the cytosol, allows determination of the topology of inner membrane proteins. Using this strategy the topology of three model proteins was correctly determined. We believe that this system could be used to predict the topology of a large number of additional proteins, especially single-spanning inner membrane proteins in E. coli. The methods for efficient protein production and topology mapping engineered during my thesis work are simple and cost-efficient and may be very valuable in future studies of membrane proteins. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 2: Manuscript.</p>

This study examined the perception of managers in the Institute of Public Administration (IPA), Riyadh, Saudi Arabia as to the way public relations is practiced within Saudi Arabia based on a comparison with Grunig's four models of public relations. Out of a population of 38 directors and director generals, 30 participated in a personal interview for a 79 percent response rate.A Q-sort analysis revealed two perceptions of public relations performance in the Institute. Perceptions differed based on educational experience, and managerial levels. Top managers with Ph.Ds tended to view performance as related to Grunig's Press Agentry and One-Way Information models. Fewer level employees saw performance related to the Two-Way Asymmetrical and Symmetrical models. The researcher concluded that the more higher educated level managers had a better understanding of the reality of performance of public relations in IPA. Main attitudes from the sorts of the participants showing a significantly and different attitude in how directors generals and directors perceived public relations was being practiced in the IPA. The directors generals and directors strongly held different views concerning the practice of public relations in the IPA. / Department of Journalism