Identification and Functional Analysis of the psaD Promoter of Chlorella vulgaris Using Heterologous Model Strains

Int J Mol Sci. 2018 Jul 6;19(7):1969. doi: 10.3390/ijms19071969.

Abstract

Chlorella has great potential as a bio-factory for production of value-added compounds. To produce the desired chemicals more efficiently in Chlorella, genetic tools for modification of Chlorella need to be developed, especially an endogenous promoter. In this study, the promoter of photosystem I protein D (psaD) from Chlorella vulgaris UTEX395 was identified. Computational analysis revealed the presence of several putative cis-acting elements, including a potential core element, and light-responsive or stress-responsive elements. Gene expression analysis in heterologous expression system in Chlamydomonasreinhardtii and Nicotianabenthamiana showed that CvpsaD promoter can be used to drive the expression of genes. Functional analysis of this promoter suggested that the initiator element (Inr) is important for its function (i.e., TATA-less promoter) and that an additional factor (e.g., downstream of the transcriptional start site) might be needed for light response. We have shown that the CvpsaD promoter is functional, but not sufficiently strong, both in microalgae and higher plant.

Keywords: Chlorella vulgaris; heterologous expression; native promoter; photosystem I protein D (psaD).

MeSH terms

  • Chlamydomonas reinhardtii / genetics
  • Chlorella vulgaris / genetics*
  • Gene Expression
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Kanamycin Kinase / genetics
  • Kanamycin Kinase / metabolism
  • Light
  • Luciferases / genetics
  • Luciferases / metabolism
  • Nicotiana / genetics
  • Photosystem I Protein Complex / genetics*
  • Plant Proteins / genetics*
  • Plants, Genetically Modified / genetics
  • Promoter Regions, Genetic / physiology*
  • Sequence Analysis, DNA
  • TATA Box

Substances

  • Photosystem I Protein Complex
  • Plant Proteins
  • Green Fluorescent Proteins
  • Luciferases
  • Kanamycin Kinase
  • Glucuronidase