A split enhanced green fluorescent protein-based reporter in yeast two-hybrid system

Kyoungsook Park, So Yeon Yi, Chang Soo Lee, Kyoon Eon Kim, Hyunsook Pai, Dai Wu Seol, Bong Hyun Chung, Moonil Kim

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We have developed a novel reporter system involving a yeast two-hybrid assay, which utilizes the reconstitution of the split EGFP reporter in order to characterize the relevant protein-protein interactions. To our knowledge, this study represents the first application of the split EGFP system as a read-out in a yeast two-hybrid assay. In comparison with the existing two-hybrid system, the bait and prey vectors were improved with regard to the reporter and the replication control element. As a result, the reconstituted EGFP has been observed to evidence a restored fluorescence upon protein-protein interactions in yeast, thereby allowing for the characterization of its interactor. The use of a split EGFP reporter has some salient advantages. Firstly, no substrates are required for the production of fluorescence. Secondly, low copy number plasmids may help to solve the protein toxicity problem, via the reduction of expression. Thirdly, this technique may prove useful in overcoming the autoactivation problem, due to the fact that the read-out of the yeast two-hybrid system is transcription-independent. Collectively, our results showed that the split EGFP reporter system might potentially be applied in yeast two-hybrid assays for the high-throughput screening of protein-protein interactions, with a simple and direct fluorescent read-out.

Original languageEnglish
Pages (from-to)107-116
Number of pages10
JournalProtein Journal
Volume26
Issue number2
DOIs
Publication statusPublished - 2007 Feb 1

Fingerprint

Two-Hybrid System Techniques
Hybrid systems
Yeast
Proteins
Assays
Fluorescence
Transcription
enhanced green fluorescent protein
Toxicity
Screening
Plasmids
Yeasts
Throughput
Substrates

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Bioengineering
  • Biochemistry
  • Organic Chemistry

Cite this

Park, Kyoungsook ; Yi, So Yeon ; Lee, Chang Soo ; Kim, Kyoon Eon ; Pai, Hyunsook ; Seol, Dai Wu ; Chung, Bong Hyun ; Kim, Moonil. / A split enhanced green fluorescent protein-based reporter in yeast two-hybrid system. In: Protein Journal. 2007 ; Vol. 26, No. 2. pp. 107-116.
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Park, K, Yi, SY, Lee, CS, Kim, KE, Pai, H, Seol, DW, Chung, BH & Kim, M 2007, 'A split enhanced green fluorescent protein-based reporter in yeast two-hybrid system', Protein Journal, vol. 26, no. 2, pp. 107-116. https://doi.org/10.1007/s10930-006-9051-2

A split enhanced green fluorescent protein-based reporter in yeast two-hybrid system. / Park, Kyoungsook; Yi, So Yeon; Lee, Chang Soo; Kim, Kyoon Eon; Pai, Hyunsook; Seol, Dai Wu; Chung, Bong Hyun; Kim, Moonil.

In: Protein Journal, Vol. 26, No. 2, 01.02.2007, p. 107-116.

Research output: Contribution to journalArticle

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AU - Chung, Bong Hyun

AU - Kim, Moonil

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