| title: | Construction of a novel bacterial two-hybrid system |
|---|---|
| reg no: | ETF5788 |
| project type: | Estonian Science Foundation research grant |
| subject: |
2.7. Biotechnology, Food and Drink Technology |
| status: | completed |
| institution: | Tartu University Institute of Technology |
| head of project: | Niilo Kaldalu |
| duration: | 01.01.2004 - 31.12.2005 |
| description: | Bacterial sigma54-dependent promoters and eukaryotic RNA polymerase II promoters are both controlled by enhancer-binding transcriptional activators. These activators have a modular structure and contain separable domains for DNA binding and transcriptional activation. Current proposal wants to utilize this similarity in design of genetic tools. The aim of this study is construction of a bacterial two-hybrid system that would become a viable alternative to the traditional yeast two-hybrid system. The method is based on activation of a sigma54-dependent promoter by a pair of chimaric proteins. In one of these hybrid proteins, the DNA-binding domain of a sigma54 ¿ specific activator is fused to a protein of interest X. The other has the transctiptional activation domain fused to protein Y. Transcription of a reporter gene will be activated only when X and Y interact. The system is designed for Escherichia coli. It will use domains of the transcriptional activator XylR of Pseudomonas putida TOL plasmid and the sigma54-dependent Pu promoter controlled by this protein. The method will be tested with several previously characterised interacting proteins and in selection of interacting partners from a cDNA expression library. |
| project group | ||||
|---|---|---|---|---|
| no | name | institution | position | |
| 1. | Niilo Kaldalu | Tartu University Institute of Technology | senior scientist | |