| 摘要 |
<p>The invention provides an improved yeast two-hybrid screening system that is optimal for high throughput screening of cDNA libraries for protein-protein interactions. The invention utilizes a combination of stably-integrated yEGFP as a reporter in the reporter yeast strain, combined with flow cytometric detection of fluorescence resulting from transcriptional activation of the reporter protein following interaction of protein-protein pairs. The system of the invention maybe conducted using liquid culture, rather than the agar plating required in conventional Y2H systems, thereby saving substantial time and resources. Moreover, by virtue of the system's liquid culture approach the invention is adaptable to high-throughput format and can be used for large-scale cDNA screening for interacting proteins with high sensitivity and low signal:noise ratios. The invention's Y2H assay/system offers convenient, quantitative and faster reporter analysis compatible with existing liquid handling robots, and also reduces labor requirements in screening cDNA libraries.</p> |
