Journal of Capital Medical University ›› 2011, Vol. 32 ›› Issue (1): 106-109.

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Construction and Identification of Yeast Two-hybrid Bait Vector pGBKT7COX-2

HE Ling-jie1, ZHANG Shu-tian2*, ZHU Sheng-tao2   

  1. 1. Department of Emergency, Beijing Friendship Hospital, Capital Medical University; 2. Department of Digestive Diseases,Beijing Friendship Hospital, Capital Medical University, Beijing Digestive Diseases Center
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-02-21 Published:2011-02-21
  • Contact: ZHANG Shu-tian

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Abstract:

Objective To construct the bait vector of cyclooxygenase-2(COX-2) in yeast two-hybrid system 3, and to test it for its autonomous activation.
Methods COX-2 gene was amplified by polymerase chain reaction(PCR) from cyclooxygenase-2(COX-2) plasmid, and the EcoRⅠ and BamHⅠ sites were introduced into it. The COX-2 gene, after sequenced, was cloned into pGBKT7. These recombinant plasmids were transferred into Y190 and they activated the reporter genes.
Results The pGBKT7-COX-2 bait vector was successfully constructed and was identified that it had no autonomous activation.
Conclusion COX-2 gene has no self-activation, bait vector could be successfully constructed, the proteins interacting with COX-2 can be detected by the yeast two-hybrid system 3.

Key words: cyclooxygenase-2, yeast two hybrid system, autonomous activation

CLC Number: