Molecular analysis of promoter elements from Propionibacterium freudenreichii

Yongzhe Piao, Nami Kawaraichi, Ryo Asegawa, Pornpimon Kiatpapan, Hisayo Ono, Mitsuo Yamashita, Yoshikatsu Murooka

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Propionibacterium freudenreichii is a commercially important microorganism that is used in the production of cheeses, cobalamin (vitamin B12), and propionic acid. Although a host-vector system in propionibacteria has been developed, there is little information available on the genetic background of the bacteria. To obtain genetic information to facilitate genetic engineering in propionibacteria, we cloned promoter regions from P. freudenreichii using Escherichia coli as a host at the first screening and a promoter-probe vector, pCVE1, which consists of the cholesterol oxidase (choA) gene from Streptomyces sp. as a reporter gene. Finally, nine clones with strong promoter activities in P. freudenreichii were screened by monitoring the choA gene expression and determining if the nucleotide sequences of the cloned DNA fragment were aligned. The initiation sites of these transcripts were determined by primer extension analysis. The putative consensus sequences corresponding to a -35 and -10 hexamer were found to be specific for P. freudenreichii, but not E. coli or other bacteria. Moreover, a new consensus heptamerous sequence between the -35 and -10 regions, termed the -16 region, was also found. It is possible that the putative consensus heptamer is functional and essential to promoter activity in P. freudenreichii. These results should provide new opportunities for controlled gene expression in P. freudenreichii.

Original languageEnglish
Pages (from-to)310-316
Number of pages7
JournalJournal of Bioscience and Bioengineering
Volume97
Issue number5
Publication statusPublished - 2004
Externally publishedYes

Fingerprint

Cholesterol Oxidase
Cholesterol
Vitamin B 12
Gene expression
Escherichia coli
Bacteria
Genes
Genetic engineering
Propionic acid
Cheeses
Vitamins
Nucleotides
Genetic Promoter Regions
Microorganisms
Screening
DNA
Monitoring
Oxidoreductases

Keywords

  • -16 region
  • Cholesterol oxidase
  • Consensus sequence
  • Promoter element
  • Propionibacterium freudenreichii

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

Cite this

Piao, Y., Kawaraichi, N., Asegawa, R., Kiatpapan, P., Ono, H., Yamashita, M., & Murooka, Y. (2004). Molecular analysis of promoter elements from Propionibacterium freudenreichii. Journal of Bioscience and Bioengineering, 97(5), 310-316.

Molecular analysis of promoter elements from Propionibacterium freudenreichii. / Piao, Yongzhe; Kawaraichi, Nami; Asegawa, Ryo; Kiatpapan, Pornpimon; Ono, Hisayo; Yamashita, Mitsuo; Murooka, Yoshikatsu.

In: Journal of Bioscience and Bioengineering, Vol. 97, No. 5, 2004, p. 310-316.

Research output: Contribution to journalArticle

Piao, Y, Kawaraichi, N, Asegawa, R, Kiatpapan, P, Ono, H, Yamashita, M & Murooka, Y 2004, 'Molecular analysis of promoter elements from Propionibacterium freudenreichii', Journal of Bioscience and Bioengineering, vol. 97, no. 5, pp. 310-316.
Piao, Yongzhe ; Kawaraichi, Nami ; Asegawa, Ryo ; Kiatpapan, Pornpimon ; Ono, Hisayo ; Yamashita, Mitsuo ; Murooka, Yoshikatsu. / Molecular analysis of promoter elements from Propionibacterium freudenreichii. In: Journal of Bioscience and Bioengineering. 2004 ; Vol. 97, No. 5. pp. 310-316.
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