Faster and more cost-effective than conventional assays

products | two- & one hybrid systems | Grow'n'Glow GFP Yeast One-Hybrid System

Two- & One Hybrid Systems

Improved sensitivity! Now available with positive controls!

Grow'n'Glow GFP Yeast One-Hybrid System

Features and Advantages:

isolates genes for novel DNA-binding proteins
maps residues and regions responsible for DNA-binding
positive colonies fluoresce bright 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

General Product Description:

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. These sort of toxic proteins would be missed during screening in a non-inducible system.

The One-Hybrid Approach with Grow'n'Glow

ORDER INFORMATION, SHIPPING & STORAGE:
order#	description	amount
GNGK03	Grow'n'Glow GFP One-Hybrid System:
	pGNG2, lyophilized DNA pJG4-5, lyophilized DNA GNGprimer 5'-PREYprimer (0.1 nmole/µl) 3'-PREYprimer (0.1 nmole/µl) yeast strain EGY48, glycerol stock prey control plasmid pJG4-5-p53 GFP positive control vector pGNG2-p53	5 µg 5 µg 500 pmole 500 pmole 500 pmole 1 ml 5 µg 5 µg
shipped on dry ice; store vectors and primers at 4 °C, yeast strain at -70 °C
GNGV02	Grow'n'Glow GFP One-Hybrid
	reporter vector pGNG2	5 µg
shipped at RT; store at 4°C;

Downloads: Handbook, pGNG2 sequence,
pJG 4 - 5 sequence

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