Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

Products

Grow'n'Glow GFP Yeast One-Hybrid System

Faster and More Cost-Effective than Conventional Assays

Features

  • 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

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. This sort of toxic proteins would be missed during screening in a non-inducible system.
The One-Hybrid Approach with Grow'n'Glow

The One-Hybrid Approach with Grow'n'Glow (to enlarge please point with mouse)

Product Amount
pGNG2, lyophilized DNA 5 µg
pJG4-5, lyophilized DNA 5 µg
GNGprimer 500 pmol
5'-PREYprimer (0.1 nmol/µl) 500 pmol
3'-PREYprimer (0.1 nmol/µl) 500 pmol
Yeast strain EGY48, glycerol stock 1 ml
Prey control plasmid pJG4-5-p53 5 µg
GFP positive control vector pGNG2-p53 5 µg

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
GNGK03 Grow 'n' Glow One-Hybrid System Kit 739,00 PDF
GNGV02 GFP reporter vector pGNG2 5 µg 439,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.

Download

  • GFP Yeast One-Hybrid System Handbook (PDF)

Vector Maps and Sequences

  • Vector map of pGNG2 (pic)
  • Vector map of pJG4-5 (pic)
  • Sequence of pGNG2 (txt)
  • Sequence of pJG4-5 (txt)