Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (9): 191-197.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0570

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Cloning and Activity Analysis of SikCDPK1 Promoter from Saussurea involucrata

SHI Guang-zhen(), WANG Zhao-ye, SUN Qi, ZHU Xin-xia()   

  1. School of Life Sciences,Shihezi University,Key Laboratory of Oasis Town and Mountain Basin System Ecology Corps,Shihezi 832003
  • Received:2022-05-09 Online:2022-09-26 Published:2022-10-11
  • Contact: ZHU Xin-xia E-mail:1106517620@qq.com;302641316@qq.com

Abstract:

The objective is to clone the SikCDPK1 promoter of Saussurea involucrata and analyze its activity, so as to lay the foundation for further understanding the transcriptional regulation mechanism of the SikCDPK1 gene in S. involucrata. TAIL-PCR was used to clone the promoter of SikCDPK1 from S. involucrata, and PlantCARE to analyze the promoter cis-acting elements. The recombinant expression vector P0∷GUS, P1∷GUS, P2∷GUS and P3∷GUS driven by full-length promoter or 5'-deletion promoter were constructed and transferred into Agrobacterium tumefaciens for transient transformation. The activities of different length promoters were analyzed by GUS staining, and the activity of GUS under low temperature and drought was determined respectively. As results, a 1 042 bp promoter sequence of SikCDPK1 was obtained. pSikCDPK1 had the core elements of eukaryotic promoter TATA-box and CAAT-box, and also contained several cis-acting elements related to stress, hormone and light response. All transformed tobacco leaves showed blue color after GUS staining, and the activity was in P0>P1>P2>P3. The GUS activity changed after low temperature and drought. In sum, the promoter of SikCDPK1 is successfully cloned and has the activity of driving downstream reporter gene expression.

Key words: Saussurea involucrata, SikCDPK1 promoter, GUS activity, transient expression