BacterioMatch II Two-Hybrid System Vector Kit is a research-use-only two-hybrid system that can be used to detect the interaction between two proteins. The system uses a reporter gene to produce a fluorescent signal, which indicates whether or not the two proteins are interacting.
The document provides instructions for the Agilent BacterioMatch II Two-Hybrid System Vector Kit (Catalog #240065), designed to characterize protein-protein interactions between proteins cloned into pBT bait and pTRG target vectors. The system utilizes an H7S3-aad4 reporter cassette in E. coli, allowing for the detection of interactions via transcriptional activation that permits growth in the presence of 3-amino-1,2,4-triazole (3-AT) and streptomycin resistance.
| pBT plasmid copy number | 5-10 copies per cell |
|---|---|
| pTRG plasmid copy number | 20-30 copies per cell |
| Reporter gene | H7S3-aad4 |
The XL1-Blue MRF' Kan strain supplied in the kit is appropriate; do not substitute another strain.
Detection is based on transcriptional activation of the H7S3 reporter gene, allowing growth in the presence of 3-amino-1,2,4-triazole (3-AT).