• Title/Summary/Keyword: formaldehyde-responsive proteins

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Isolation of formaldehyde-responsive proteins in Arabidopsis (Formaldehyde에 반응하는 애기장대 단백질의 분리)

  • Kwon, Mi;Park, Hyun Jin;Seo, Jae Hyun
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.4
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    • pp.52-60
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    • 2007
  • Plant can detoxify the effect of the volatile organic compounds (VOC) such as formaldehyde and toluene, however, mechanisms of VOC detoxification are largely unknown in plant system. This study was performed to investigate phenotypic changes of Arabidopsis seedlings upon treatment of either formalin or toluene. Formalin treatment up to twenty four hours didn't cause any significant phenotypic damages on the leaf surface of 27 DAG Arabidopsis seedlings. However, the protein profile of formalin-treated seedlings was significantly different from that of mock control. Using automated electrophoresis system, the molecular weight of each formaldehyde-responsive protein (FRP) was predicted and its formaldehyde-dependent expression was confirmed at transcription level by quantitative real-time RT-PCR analysis. Four FRPs isolated in this study are the novel proteins with unknown functions but highly homologous to the stress-related proteins.

Isolation of Putative in vivo Hoxc8 Downstream Target Genes Using ChIP-Cloning Method

  • Chung, Hyun-Joo;Kang, Myeng-Mo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.14 no.1
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    • pp.47-53
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    • 2008
  • Hox genes are known to be transcription factors controlling vertebrate pattern formation along the anteroposterior body axis by regulating many target gene expressions during vertebrate embryogenesis. In order to isolate in vivo Hox responsive target genes, ChIP-cloning technique has been applied using Hoxc8 antibody. Here murine embryo of day 11.5 post coitum (E11.5) highly expressing Hoxc8 gene was used after removing head and tail portions where Hoxc8 is rarely expressing. After fixation with formaldehyde, the chromatin DNAs harboring bound proteins were isolated. After sonication, about 0.5- to 1 Kb chromatin DNAs were immunoprecipitated with anti Hoxc8 antibody. After removing the bound proteins with proteinase K, DNAs were isolated, cloned into the pBluescsript II SK vector, and then sequenced. Total 33 random clones sequenced were anlalyzed to be located at 12 different genomic regions. Among these, 8 turned out to be introns and 4 were intergenic regions localized in random chromosomes. The base composition of total cloned genomic sequences (6608 bp) were AT-rich, i.e., 40% GC. When the Hoxc8 core binding sites, such as TAAT, ATTA, TTAT, and ATAA were analyzed total number of 55, 45, 54, and 55 were found, respectively, which are than twice as many as expected number of 26. Although this in silico analysis does not mean that the ChIP-cloned sequence is real Hoxc8 regulatory element in vivo, these results strongly imply that the DNA fragments cloned through chromatin immunoprecipitation could be very much likely the putative Hoxc8 downstream target genes.

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