Bacteriophage lambda replication and phage display

点击翻译按钮获取中文摘要

Our research program involves fundamental studies on bacteriophage lambda virus replication, and genetic engineering attempts to develop a platform for making multiplication-defective lambda virus particles displaying gene fusions. These viral particles can have utility as vaccines and for gene therapy. The platform accomplishes the divergent tasks of enabling very high concentration production of the viral particles, yet effectively killing the virus by introducing genetic modifications that prevent it from multiplying in animal cells or in the environment. The platform will simultaneously produce encapsulated viral DNA and gene-fusion display particles. The gene-fusions displayed on the viral particles enable them to recognize target proteins on specific animal cells, enhancing recognition and viral uptake. Once in the cell, the gene expression cassette that was inserted into the viral genome will encode for the synthesis of a protein(s) that can induce a vaccine response, or support gene therapy. Dr. Hayes has successfully designed gene fusion phage display vaccines to PorcinePCV2 and Bovine BVDV2 viruses, to Ebola and Marburg viruses, and several to anthrax. Advantages of symbiotic research goals: Many of the platform technology gene expression vectors have proven of immense value to our gene expression and viral replication studies. Here, we try to understand how replication initiation from the lambda virus is silenced, which we found involves the participation of a micro RNA. We have identified lambda mutations that escape silencing (Sip) and seek to characterize their effect. Studies also focus on the activities of the lambda replication initiation genes O and P, and host cell proteins. Our particular focus is the interaction between the host DnaB helicase required to unwind DNA during replication and P, which is very toxic to the cell, and appears to stimulate a very high frequency of chromosomal mutation. Lambda escapes the effects of P. We seek to characterize the effects of P on host cell metabolism, and the regulation of gene P expression and P activity in cell.