Expression and Construction of Yeast Expression Vector Containing CsTCTP1 Gene from Cucumber in Yeast Two-Hybrid System
Qiu-Min Chen1, Na Cui1, 2, *, Yang Yu1, Xiang-Nan Meng1, Hai-Yan Fan1, 2, *
1 College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110866, PR China
2 Key Laboratory of Protected Horticulture of Ministry of Education, Shenyang Agricultural University, Shenyang 110866, PR China
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.
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.
Specific PCR, conventional sequencing, heat shock conversion method and TE/LiAC transformation method.
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.
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.
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