Expression and Construction of Yeast Expression Vector Containing CsTCTP1 Gene from Cucumber in Yeast Two-Hybrid System

Abstract

Background: The translationally controlled tumor protein (TCTP) was originally found in tumor tissue, and later found in other tissues. Initially, TCTP was considered a kind of growth-associated protein. Recent studies have shown that TCTP has many biological functions. Objective: To verification of CsTCTP1 gene function by yeast two-hybrid system, the pGBKT7- CsTCTP1 yeast expression vector was constructed and cytotoxicity and self-activating activity were detected, which could lay the foundation for further studies on gene function and make a preparation for verification of CsTCTP1 gene function by yeast two-hybrid system. Method: Specific PCR, conventional sequencing, heat shock conversion method and TE/LiAC transformation method. Results: We constructed a yeast expression vector containing the CsTCTP1 gene. The CsTCTP1 coding sequence was inserted into a pGBKT7 vector as a bait protein and then transformed into the Y2HGold yeast stain. Conclusion: We found that CsTCTP1 protein had no cytotoxic effect and could not be self-activated. The constructed bait expression vector can be used in the subsequent yeast two - hybrid detection system.