Grow'n'Glow GFP Yeast One-Hybrid System
Faster and More Cost-Effective than Conventional Assays
- Isolates genes for novel DNA-binding proteins
- Maps residues and regions responsible for DNA-binding
- Positive colonies fluoresce brightly green
- Therefore, one-step selection of potential positives under UV light
- No time-consuming beta-galactosidase lift assays as essential in most conventional assays
- Finds potentially toxic proteins by screening with galactose-inducible expression libraries
- The Grow'n'Glow GFP One-Hybrid kit isolates genes for proteins that bind a specific DNA element of interest. In addition to finding novel DNA-binding proteins,
the one-hybrid system can be used to investigate the bases and amino acids involved in specific DNA-protein interactions. Proteins can be found that bind to any
short DNA element of interest. The Grow'n'Glow system offers maximal sensitivity because detection of the DNA-protein interaction occurs in vivo,
where proteins are more likely to be in their native conformation.
- Use of the Green Fluorescent Protein (GFP) reporter eliminates the time-consuming beta-galactosidase lift assays. The particular GFP variant expressed by the
GFP reporter plasmid, GFPuv, has the same excitation and emission maxima as wild-type GFP, but is 18 times brighter than the wild-type variant. Positive colonies
with DNA-binding proteins glow brightly green and can be easily detected by placing the plate with the yeast colonies on a standard 300 nm UV transilluminator
or under a UV lamp in a darkroom.
- Expression of fusion proteins by the prey plasmid is controlled by the GAL1 upstream-activation sequence. In yeast with an intact galactose regulatory system,
the GAL1 activation sequence is induced by galactose and repressed by glucose. This regulation inhibits expression of library/activation domain fusions until
the actual screening, which prevents any potentially toxic library proteins affecting growth of the yeast. This sort of toxic proteins would be missed during
screening in a non-inducible system.
The One-Hybrid Approach with Grow'n'Glow (to enlarge please point with mouse)
|pGNG2, lyophilized DNA
|pJG4-5, lyophilized DNA
|5'-PREYprimer (0.1 nmol/µl)
|3'-PREYprimer (0.1 nmol/µl)
|Yeast strain EGY48, glycerol stock
|Prey control plasmid pJG4-5-p53
|GFP positive control vector pGNG2-p53
- For shipping and storage information please click on Order#.
||Grow 'n' Glow One-Hybrid System
||GFP reporter vector pGNG2
- All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.
- GFP Yeast One-Hybrid System Handbook (PDF)
Vector Maps and Sequences
- Vector map of pGNG2 (pic)
- Vector map of pJG4-5 (pic)