Improvement of the bioluminescence reporter system for real-time monitoring of circadian rhythms in the cyanobacterium Synechocystis sp. strain PCC 6803

• Kucho Ken-ichi

  Center for Gene Research, Nagoya University

• Aoki Kazuhiro

  Division of Material Science, Graduate School of Science, Nagoya University

• Itoh Shigeru

  Division of Material Science, Graduate School of Science, Nagoya University

• Ishiura Masahiro

  Center for Gene Research, Nagoya University Division of Biological Science, Graduate School of Science, Nagoya University Bio-oriented Technology Research Advancement Institution (BRAIN), Nagoya University

抄録

Circadian rhythm is a self-sustaining oscillation whose period length coincides with the 24-hour day-night cycle. A powerful tool for circadian clock research is the real-time automated bioluminescence monitoring system in which a promoter region of a clock-controlled gene is fused to a luciferase reporter gene and rhythmic regulation of the promoter activity is monitored as bioluminescence. In the present study, we greatly improved the bioluminescence reporter system in the cyanobacterium Synechocystis sp. strain PCC 6803. We fused an 805-bp promoter region of the dnaK gene seamlessly to the luxA coding sequence and inte-grated the P_dnaK::luxAB fusion gene into a specific intergenic region of the Syn-echocystis genome (targeting site 1). The resulting new reporter strain, PdnaK::luxAB(–), showed 12 times the bioluminescence intensity of the standard reporter strain, CFC2. Furthermore, we generated strain PdnaK::luxAB(+), in which the P_dnaK::luxAB fusion gene and the selection-marker spectinomycin resistance gene are transcribed in opposite directions. The PdnaK::luxAB(+) strain showed 19 times the bioluminescence intensity of strain CFC2. The procedures used to increase the bioluminescence intensity are especially useful for bioluminescence monitoring of genes with low promoter activity. In addition, these reporter constructs facilitate bioluminescence monitoring of any gene because the promoter fragments they contain can easily be replaced by digestion with unique restriction enzymes. They would therefore contribute to a genome-wide analysis of gene expression in Synechocystis.<br>

収録刊行物

• Genes & Genetic Systems

  Genes & Genetic Systems 80 (1), 19-23, 2005

  日本遺伝学会

キーワード

• bioluminescence
• circadian rhythm
• <i>dnaK</i>
• <i>luxAB</i>
• <i>Synechocystis</i>

詳細情報

• CRID: 1390282680450672128
• NII論文ID: 10015512154
• NII書誌ID: AA11077421
• DOI: 10.1266/ggs.80.19
• ISSN: 18805779
• ISSN: 13417568
• NDL書誌ID: 7299555
• PubMed: 15824452
• Web Site: http://id.ndl.go.jp/bib/7299555
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7299555
• Web Site: http://www.jstage.jst.go.jp/article/ggs/80/1/80_1_19/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可