Expression of different fragments of integrin beta 3 subunit in the yeast two-hybrid system and their activation of reporter gene

Jian Bo Yang, Jia Yao, Jie Yang

Research output: Contribution to journalArticle

Abstract

Aim: To construct the bait vectors containing different amino terminal fragments of integnin beta 3 subunit in the yeast two-hybrid system, and to test whether their expressed product could affect the growth of yeast cells and activate the transcription of reporter genes. Methods: The cDNA fragments encoding different sizes of integrin beta 3 amino terminal region were amplified by PCR, then cloned into pAS2-1. These fragments included A1 (55E-199Q), A2(235K-385S), A3 (4 I-454T), A4 (4 I-77S), A5 (4 I-147S) and A6 (4 I-284S). After verified by sequencing, these bait vectors were transferred into the yeast two-hybrid host Y190 to assay whether the transcription of the reporter genes was activated. Results: The bait vectors of different sizes of integnin beta 3 amino terminal region were constructed successfully. It was found that all fragments were not toxic to Y190. But fragment 4 I-77S and another mutant fragment (4 I-F56 + Q) activated the expression of the reporter genes. Conclusion: Fragments A1, A2 and A3 could be used as the bait in the yeast two-hybrid system. The amino terminal fragment (4 I-77S) can activate the transcription of the Gal4-dependent reporter genes when it is fused to the Gal4 DNA binding domain.

Original languageEnglish (US)
Pages (from-to)584-589
Number of pages6
JournalJournal of China Pharmaceutical University
Volume36
Issue number6
StatePublished - Dec 1 2005

Keywords

  • Integrin alpha II b/beta 3
  • Transcriptional activition
  • Yeast two-hybrid

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Fingerprint Dive into the research topics of 'Expression of different fragments of integrin beta 3 subunit in the yeast two-hybrid system and their activation of reporter gene'. Together they form a unique fingerprint.

  • Cite this