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Titolo:
ANALYSIS OF ACTINOMYCETE COMMUNITIES BY SPECIFIC AMPLIFICATION OF GENES ENCODING 16S RIBOSOMAL-RNA AND GEL-ELECTROPHORETIC SEPARATION IN DENATURING GRADIENTS
Autore:
HEUER H; KRSEK M; BAKER P; SMALLA K; WELLINGTON EMH;
Indirizzi:
BIOL BUNDESANSTALT LAND & FORSTWIRTSCHAFT,INST BIOCHEM & PFLANZENVIROL,MESSEWEG 11-12 D-38104 BRAUNSCHWEIG GERMANY BIOL BUNDESANSTALT LAND & FORSTWIRTSCHAFT,INST BIOCHEM & PFLANZENVIROL D-38104 BRAUNSCHWEIG GERMANY UNIV WARWICK,DEPT BIOL SCI COVENTRY CV4 7AL W MIDLANDS ENGLAND
Titolo Testata:
Applied and environmental microbiology
fascicolo: 8, volume: 63, anno: 1997,
pagine: 3233 - 3241
SICI:
0099-2240(1997)63:8<3233:AOACBS>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE CHAIN-REACTION; RIBOSOMAL-RNA; DNA AMPLIFICATION; ESCHERICHIA-COLI; NONSTERILE SOIL; NUCLEIC-ACIDS; GC-CLAMP; FRAGMENTS; BACTERIA; PLASMID;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
60
Recensione:
Indirizzi per estratti:
Citazione:
H. Heuer et al., "ANALYSIS OF ACTINOMYCETE COMMUNITIES BY SPECIFIC AMPLIFICATION OF GENES ENCODING 16S RIBOSOMAL-RNA AND GEL-ELECTROPHORETIC SEPARATION IN DENATURING GRADIENTS", Applied and environmental microbiology, 63(8), 1997, pp. 3233-3241

Abstract

A group-specific primer, F243 (positions 226 to 243, Escherichia colinumbering), was developed by comparison of sequences of genes encoding 16S rRNA (16S rDNA) for the detection of actinomycetes in the environment with PCR and temperature or denaturing gradient gel electrophoresis (TGGE or DGGE, respectively). The specificity of the forward primer in combination with different reverse ones was tested with genomic DNA from a variety of bacterial strains. Most actinomycetes investigated could be separated by TGGE and DGGE, with both techniques giving similar results. Two strategies were employed to study natural microbial communities. First, we used the selective amplification of actinomycete sequences (E. coli positions 226 to 528) for direct analysis of the products in denaturing gradients. Second, a nested PCR providing actinomycete-specific fragments (E. coli positions 226 to 1401) was used which served as template for a PCR when conserved primers were used. Theproducts (E. coli positions 968 to 1401) of this indirect approach were then separated by use of gradient gels. Both approaches allowed detection of actinomycete communities in soil. The second strategy allowed the estimation of the relative abundance of actinomycetes within thebacterial community. Mixtures of PCR-derived 16S rDNA fragments were used as model communities consisting of five actinomycetes and five other bacterial species. Actinomycete products were obtained over a 100-fold dilution range of the actinomycete DNA in the model community by specific PCR; detection of the diluted actinomycete DNA was not possible when conserved primers were used. The methods tested for detection were applied to monitor actinomycete community changes in potato rhizosphere and to investigate actinomycete diversity in different soils.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 07/07/20 alle ore 06:23:04