Expression of bkt and bch genes from Haematococcus pluvialis in transgenic Chlamydomonas

Sci China Life Sci. 2014 Oct;57(10):1028-33. doi: 10.1007/s11427-014-4729-8. Epub 2014 Sep 9.

Abstract

β-carotene ketolase and β-carotene hydroxylase encoded by bkt and bch, respectively, are key enzymes required for astaxanthin biosynthesis in Haematococcu pluvialis 34-1n. Two expression vectors containing cDNA sequences of bkt and bch were constructed and co-transformed into cell-wall-deficient Chlamydomonas reinhardtii CC-849. Transgenic algae were screened on TAP agar plates containing 10 μg mL(-1) Zeomycin. PCR-Southern analysis showed that bkt and bch were integrated into the genomes of C. reinhardtii. Transcripts of bkt and bch were further confirmed by RT-PCR-Southern analysis. Compared with the wild type, transgenic algae produced 29.04% and 30.27% more carotenoids and xanthophylls, respectively. Moreover, the transgenic algae could accumulate 34% more astaxanthin than wild type. These results indicate that foreign bkt and bch genes were successfully translated into β-carotene ketolase and β-carotene hydroxylase, which were responsible for catalyzing the biosynthesis of astaxanthin in transgenic algae.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algal Proteins / genetics
  • Algal Proteins / metabolism*
  • Carotenoids / chemistry
  • Cell Nucleus / metabolism
  • Cell Wall
  • Chlamydomonas / genetics*
  • Chlorophyta / enzymology*
  • Chlorophyta / genetics
  • DNA, Complementary
  • Genetic Techniques
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism*
  • Oxygenases / genetics
  • Oxygenases / metabolism*
  • Transgenes
  • Xanthophylls / chemistry

Substances

  • Algal Proteins
  • DNA, Complementary
  • Xanthophylls
  • Carotenoids
  • astaxanthine
  • Mixed Function Oxygenases
  • Oxygenases
  • beta-carotene ketolase
  • beta-carotene hydroxylase