Transcriptome Analysis of Bacillus subtilis by DNA Microarray Technique

  • Kang, Choong-Min (Department of Food Science, Youngdong University, Department of Biotechnology, Fukuyama University) ;
  • Yoshida, Ken-Ichi (Department of Biotechnology, Fukuyama University) ;
  • Matsunaga, Masayuki (Department of Biotechnology, Fukuyama University) ;
  • Kobayashi, Kazuo (Graduate School of Biological Science, Nara Institute of Science and Technology (NAIST)) ;
  • Ueda, Minoru (Biomedical Group, Takara Shuzo Co.) ;
  • Ogasawara, Naotake (Graduate School of Biological Science, Nara Institute of Science and Technology (NAIST)) ;
  • Fujita, Yasutaro (Department of Biotechnology, Fukuyama University)
  • 발행 : 2000.06.23

초록

The complete genome sequence of a Gram-positive bacterium .Bacillus subtilis has recently been reported and it is now clear that more than 50% of its ORFs have no known function (1). To study the global gene expression in B. subtilis at single gene resolution, we have tested the glass DNA microarrays in a step-wise fashion. As a preliminary experiment, we have created arrays of PCR products for 14 ORF whose transcription patterns have been well established through transcriptional mapping analysis. We measured changes in mRNA transcript levels between early exponential and stationary phase by hybridizing fluorescently labeled cDNA (with Cy3-UTP and Cy5-UTP) onto the array. We then compared the microarray data to confirm that the transcription patterns of these genes are well consistent with the known Northern analysis data. Since the preliminary test has been successful, we scaled up the experiments to ${\sim}$94% of the 4,100 annotated ORFs for the complete genome sequence of B. subtilis. Using this whole genomic microarray, we searched genes that are catabolite-repressive and those that are under the control of ${\sigma}^{Y}$, one of the functionally unknown ECF sigma factors. From these results, we here report that we have established DNA microarray techniques that are applicable for the whole genome of B. subtilis.

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