2012 Annual Report 1a.Objectives (from AD-416): To develop and test models for predicting and controlling translational efficiency of gene expression, and to design and carry out experiments to validate model predictions. 1b.Approach (from AD-416): Computer models will be developed using free energy of ribosome binding as a factor in translational efficiency. Models will be tested with collaborators using recombinant plasmids in Escherichia coli. 3.Progress Report: This project is related to in-house objective 2, to understand how bacterial metabolism works at a fundamental level, will increase our ability to improve probiotic bacterial survival in acid solutions. The central paradigm of gene expression in molecular biology states that deoxyribonucleic acid (DNA) makes ribonucleic acid (RNA) which makes protein. When expressing foreign genes in Escherichia coli and other microorganisms, there have been repeated difficulties with the RNA to protein step, which is called translation. This research directly addresses this bottleneck. A novel computer model for how translation efficiency can be improved has been developed in collaboration with researchers at NCSU’s Computer Science Department and the Department of Microbiology at NCSU. Research that has been done to validate the computer model includes: measuring ribosome binding to messenger RNA and how modified genes are translated. It has been shown that modifications based on the computer model significantly improve translational efficiency. This research may lead to a breakthrough in getting genes that are difficult to express in Escherichia coli and other industrial strains of bacteria to produce needed proteins.