Chromosomal integration of a binding domain:bait gene into yeast enhances detection in the two-hybrid system

Shuhei Miyashita, Yukio Shirako

研究成果: Article査読

4 被引用数 (Scopus)

抄録

We have developed a modified yeast two-hybrid system using the GAL4 transcription activator by integrating a BD:bait gene (GAL4 binding domain:bait gene) into the host chromosome. Locus-specific integration by homologous recombination and use of a strong transcription promoter enabled uniform expression of the integrated BD:bait gene in all host cells. Moreover, after introduction of an AD:prey (GAL4 activation domain:prey) plasmid, false positives were nearly undetectable and false negatives were decreased significantly. Using this improved method a cDNA library could be efficiently screened using roughly 1/10 of the materials required for conventional sequential two-hybrid screen.

本文言語English
ページ(範囲)179-184
ページ数6
ジャーナルJournal of Microbiological Methods
73
2
DOI
出版ステータスPublished - 2008 5月

ASJC Scopus subject areas

  • 微生物学
  • 分子生物学
  • 微生物学(医療)

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