Isolation and characterization of a 3-hydroxy-3-methylglutaryl coenzyme A reductase 2 promoter from Salvia miltiorrhiza

  • Piotr Szymczyk
  • Renata Grąbkowska
  • Ewa Skała
  • Marta Żebrowska
  • Ewa Balcerczak
  • Agnieszka Jeleń
Original Article


The aim of the present study was to isolate and characterize the 5′ regulatory region of Salvia miltiorrhiza 3-hydroxy-3-methylglutaryl coenzyme A reductase 2 gene. The entire fragment is 2696 bp long and consists of the promoter, 5′UTR and 85 nucleotide 5′ fragments of the CDS region. The results of in silico bioinformatic tests indicate that the promoter region contains repetitions of many potential cis-active elements serving as the recognition sites for transcription factors. Data obtained from the in silico tests are verified by co-expression studies based on A. thaliana microarray data to make them more probably to occur. The bioinformatic analysis indicated no tandem repeats or CpNpG islands in the promoter. However, potential target sites for miRNA 156 and miRNA 159 were found in the 5′ UTR segment. In addition, two possible polymorphic sites, A2719G and A2744C, were found in the CDS region. Finally, the activity of isolated fragment was evaluated experimentally by quantitative RT–PCR. The promoter activity of the isolated 2696 bp HMGR2 gene fragment was confirmed by RT–PCR studies of in vitro cultured, transformed S. miltiorrhiza plants. Analysis of the RT–PCR results revealed temporal changes in the promoter activity occurring in response to treatment by five abiotic factors: auxin, gibberellin, salicylic acid, abscisic acid and 100 mM NaCl.


miRNA Polymorphic site Promoter RT–PCR Trans-factor 



Abscisic acid


Coding DNA sequence




Gibberellic acid


3-hydroxy-3-methylglutaryl coenzyme A reductase




Methyl jasmonate


Ser/Thr protein phosphatase 2A


5′ untranslated region



Authors are extremely grateful to Dean Elżbieta Mikiciuk-Olasik, the former Dean of the Faculty of Pharmacy, Medical University of Łódź, and Dean Daria Orszulak-Michalak for providing financial support. Authors gratefully acknowledge the technical assistance of Wacław Prószyński.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Supplementary material

13562_2017_434_MOESM1_ESM.docx (2.2 mb)
Supplementary material 1 (DOCX 2207 kb)


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Copyright information

© Society for Plant Biochemistry and Biotechnology 2017

Authors and Affiliations

  • Piotr Szymczyk
    • 1
  • Renata Grąbkowska
    • 2
  • Ewa Skała
    • 2
  • Marta Żebrowska
    • 3
  • Ewa Balcerczak
    • 3
  • Agnieszka Jeleń
    • 3
  1. 1.Department of Pharmaceutical BiotechnologyMedical University of ŁódźLodzPoland
  2. 2.Department of Biology and Pharmaceutical BotanyMedical University of ŁódźLodzPoland
  3. 3.Department of Pharmaceutical Biochemistry and Molecular DiagnosticsMedical University of ŁódźLodzPoland

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