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Titolo:
Design of PCR primers and a gene probe for extensive detection of poly(3-hydroxybutyrate) (PHB)-degrading bacteria possessing fibronectin type III linker type-PHB depolymerases
Autore:
Sei, K; Nakao, M; Mori, K; Ike, M; Kohno, T; Fujita, M;
Indirizzi:
Yamanashi Univ, Fac Engn, Dept Civil & Environm Engn, Kofu, Yamanashi 4008511, Japan Yamanashi Univ Kofu Yamanashi Japan 4008511 ofu, Yamanashi 4008511, Japan Osaka Univ, Grad Sch Engn, Dept Environm Engn, Suita, Osaka 5650871, JapanOsaka Univ Suita Osaka Japan 5650871 nm Engn, Suita, Osaka 5650871, Japan
Titolo Testata:
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
fascicolo: 6, volume: 55, anno: 2001,
pagine: 801 - 806
SICI:
0175-7598(200106)55:6<801:DOPPAA>2.0.ZU;2-T
Fonte:
ISI
Lingua:
ENG
Soggetto:
BETA-HYDROXYBUTYRATE METABOLISM; RIBOSOMAL-RNA; POPULATIONS; DEGRADATION; SOIL;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
20
Recensione:
Indirizzi per estratti:
Indirizzo: Sei, K Yamanashi Univ, Fac Engn, Dept Civil & Environm Engn, 4-3-11 Takeda, Kofu,Yamanashi 4008511, Japan Yamanashi Univ 4-3-11 Takeda Kofu YamanashiJapan 4008511 11, Japan
Citazione:
K. Sei et al., "Design of PCR primers and a gene probe for extensive detection of poly(3-hydroxybutyrate) (PHB)-degrading bacteria possessing fibronectin type III linker type-PHB depolymerases", APPL MICR B, 55(6), 2001, pp. 801-806

Abstract

For rapid and sensitive detection of poly(3-hydroxybutyrate) (PHB)-degrading bacteria, a PCR primer set (PHB primers) and a gene probe (PHB probe) were designed, based on the homologous regions of six fibronectin type III linker domain-encoding sequences laid on a variety of PHB depolymerase genes listed in the GenBank. PCR using PHB primers amplified DNA fragments with the expected sizes from all the tested bacterial strains used for primer design: and all of the amplified fragments gave positive signals by Southern hybridization with the PHB probe. No amplified fragments were observed from negative controls. To evaluate the availability of the PHB primers and PHB probe, they were applied to 57 wild-type, PHB-degrading bacteria newly isolated from a variety of environments. The PHB primers amplified DNA fragments with expected sizes from 50 of the 57 wild-type strains, while the PHB probe showed positive signals against the amplified fragments from 47 strains. These results suggest that the primer and probe system established in this study can detect a considerable proportion of the potential PHB-degradingbacteria and can be applied to evaluate PHB-degradation potential in a natural environment, in combination with direct DNA extraction methods.

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Documento generato il 30/03/20 alle ore 17:55:53