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Extraction and detection of guanosine 5'-diphosphate-3'-diphosphate in amino acid starvation cells of Clavibacter michiganensis.
Bai, Kaihong; Chen, Xing; Jiang, Na; Lyu, Qingyang; Li, Jianqiang; Luo, Laixin.
Afiliação
  • Bai K; Beijing Key Laboratory of Seed Disease Testing and Control, Department of Plant Pathology, China Agricultural University, Beijing, 100193, People's Republic of China.
  • Chen X; Beijing Key Laboratory of Seed Disease Testing and Control, Department of Plant Pathology, China Agricultural University, Beijing, 100193, People's Republic of China.
  • Jiang N; Beijing Key Laboratory of Seed Disease Testing and Control, Department of Plant Pathology, China Agricultural University, Beijing, 100193, People's Republic of China.
  • Lyu Q; Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
  • Li J; Beijing Key Laboratory of Seed Disease Testing and Control, Department of Plant Pathology, China Agricultural University, Beijing, 100193, People's Republic of China.
  • Luo L; Beijing Key Laboratory of Seed Disease Testing and Control, Department of Plant Pathology, China Agricultural University, Beijing, 100193, People's Republic of China. luolaixin@cau.edu.cn.
Braz J Microbiol ; 52(3): 1573-1580, 2021 Sep.
Article em En | MEDLINE | ID: mdl-33837930
Guanosine 5'-diphosphate-3'-diphosphate (ppGpp) is a small molecule nucleotide alarmone that can accumulate under the amino acid starvation state and trigger the stringent response. This study reported the extraction of ppGpp from the Gram-positive bacteria Clavibacter michiganensis through methods using formic acid, lysozyme, or methanol. Following extraction, ppGpp was detected through ultra-high-performance liquid chromatography (UHPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The methanol method showed the highest extraction efficiency for ppGpp among the three methods tested. C. michiganensis cells in exponential growth phase was induced in amino acid starvation by serine hydroxamate (SHX) and used for ppGpp extraction and detection. When using the methanol extraction method, the results showed that ppGpp concentrations in SHX-treated samples were 15.645 nM, 17.656 nM, 20.372 nM, and 19.280 nM at 0 min, 15 min, 30 min and 1 h, respectively, when detected using LC-MS/MS. This is the first report on ppGpp extraction and detection in Clavibacter providing a new idea and approach for nucleotide detection and extraction in bacteria.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Difosfatos / Guanosina Tetrafosfato Tipo de estudo: Diagnostic_studies Idioma: En Revista: Braz J Microbiol Ano de publicação: 2021 Tipo de documento: Article País de publicação: Brasil
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Difosfatos / Guanosina Tetrafosfato Tipo de estudo: Diagnostic_studies Idioma: En Revista: Braz J Microbiol Ano de publicação: 2021 Tipo de documento: Article País de publicação: Brasil