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One- and Two-Hybrid Systems

GNGK03 - Grow'n'Glow GFP One-Hybrid System

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Starting at: $1,269.00

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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

1-hybrid

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.
prey
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.

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