The Open Plant Science Journal
2017, 10 : 55-61Published online 2017 June 30. DOI: 10.2174/1874294701710010055
Publisher ID: TOPSJ-10-55
RESEARCH ARTICLE
Expression and Construction of Yeast Expression Vector Containing Gene from Cucumber in Yeast Two-Hybrid System
* Address correspondence to this authors at the College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110866, PR China Tel: +86 2488487163, Fax: +86 2488487163; E-mails: ; syaua@163.com
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.