PACE analysis of the heteroduplexes formed between PCR-amplified 16S rRNA genes: Estimation of sequence similarity and rDNA complexity

Romilio T. Espejo, Carmen Gloria Feijóo, Jaime Romero, Mónica Vásquez

Resultado de la investigación: Article

49 Citas (Scopus)

Resumen

Analysis of the 16S rRNA genes retrieved directly from different environments has proven to be a powerful tool that has greatly expanded our knowledge of microbial diversity and phylogeny. It is shown here that sequence similarity between 80 and 100% among 16S rDNAs can be estimated by the electrophoretic migration of their heteroduplexes. This was measured by hybridization and electrophoresis in polyacrylamide gels of the product obtained after PCR amplification of almost the entire 16S rRNA gene from different bacterial species. These heteroduplexes were also observed after amplification of samples containing DNA from two or more bacterial species and a procedure was applied to identify reliably heteroduplexes among the amplification products. The electrophoretic migration of the heteroduplexes observed after PCR was used to detect the presence of 16S rDNAs with different sequences in DNA extracted from both a mixture of two bacterial species and samples containing a natural bacterial community.

Idioma originalEnglish
Páginas (desde-hasta)1611-1617
Número de páginas7
PublicaciónMicrobiology
Volumen144
N.º6
DOI
EstadoPublished - 1 ene 1998

Huella dactilar

Heteroduplex Analysis
Ribosomal DNA
rRNA Genes
Polymerase Chain Reaction
Phylogeny
Polyacrylamide Gel Electrophoresis
DNA

ASJC Scopus subject areas

  • Microbiology

Citar esto

@article{87f8454fac194f658a12865a579d78b1,
title = "PACE analysis of the heteroduplexes formed between PCR-amplified 16S rRNA genes: Estimation of sequence similarity and rDNA complexity",
abstract = "Analysis of the 16S rRNA genes retrieved directly from different environments has proven to be a powerful tool that has greatly expanded our knowledge of microbial diversity and phylogeny. It is shown here that sequence similarity between 80 and 100{\%} among 16S rDNAs can be estimated by the electrophoretic migration of their heteroduplexes. This was measured by hybridization and electrophoresis in polyacrylamide gels of the product obtained after PCR amplification of almost the entire 16S rRNA gene from different bacterial species. These heteroduplexes were also observed after amplification of samples containing DNA from two or more bacterial species and a procedure was applied to identify reliably heteroduplexes among the amplification products. The electrophoretic migration of the heteroduplexes observed after PCR was used to detect the presence of 16S rDNAs with different sequences in DNA extracted from both a mixture of two bacterial species and samples containing a natural bacterial community.",
keywords = "16s rRNA, Diversity, Heteroduplex, Phylogeny",
author = "Espejo, {Romilio T.} and Feij{\'o}o, {Carmen Gloria} and Jaime Romero and M{\'o}nica V{\'a}squez",
year = "1998",
month = "1",
day = "1",
doi = "10.1099/00221287-144-6-1611",
language = "English",
volume = "144",
pages = "1611--1617",
journal = "Microbiology (United Kingdom)",
issn = "1350-0872",
publisher = "Society for General Microbiology",
number = "6",

}

PACE analysis of the heteroduplexes formed between PCR-amplified 16S rRNA genes : Estimation of sequence similarity and rDNA complexity. / Espejo, Romilio T.; Feijóo, Carmen Gloria; Romero, Jaime; Vásquez, Mónica.

En: Microbiology, Vol. 144, N.º 6, 01.01.1998, p. 1611-1617.

Resultado de la investigación: Article

TY - JOUR

T1 - PACE analysis of the heteroduplexes formed between PCR-amplified 16S rRNA genes

T2 - Estimation of sequence similarity and rDNA complexity

AU - Espejo, Romilio T.

AU - Feijóo, Carmen Gloria

AU - Romero, Jaime

AU - Vásquez, Mónica

PY - 1998/1/1

Y1 - 1998/1/1

N2 - Analysis of the 16S rRNA genes retrieved directly from different environments has proven to be a powerful tool that has greatly expanded our knowledge of microbial diversity and phylogeny. It is shown here that sequence similarity between 80 and 100% among 16S rDNAs can be estimated by the electrophoretic migration of their heteroduplexes. This was measured by hybridization and electrophoresis in polyacrylamide gels of the product obtained after PCR amplification of almost the entire 16S rRNA gene from different bacterial species. These heteroduplexes were also observed after amplification of samples containing DNA from two or more bacterial species and a procedure was applied to identify reliably heteroduplexes among the amplification products. The electrophoretic migration of the heteroduplexes observed after PCR was used to detect the presence of 16S rDNAs with different sequences in DNA extracted from both a mixture of two bacterial species and samples containing a natural bacterial community.

AB - Analysis of the 16S rRNA genes retrieved directly from different environments has proven to be a powerful tool that has greatly expanded our knowledge of microbial diversity and phylogeny. It is shown here that sequence similarity between 80 and 100% among 16S rDNAs can be estimated by the electrophoretic migration of their heteroduplexes. This was measured by hybridization and electrophoresis in polyacrylamide gels of the product obtained after PCR amplification of almost the entire 16S rRNA gene from different bacterial species. These heteroduplexes were also observed after amplification of samples containing DNA from two or more bacterial species and a procedure was applied to identify reliably heteroduplexes among the amplification products. The electrophoretic migration of the heteroduplexes observed after PCR was used to detect the presence of 16S rDNAs with different sequences in DNA extracted from both a mixture of two bacterial species and samples containing a natural bacterial community.

KW - 16s rRNA

KW - Diversity

KW - Heteroduplex

KW - Phylogeny

UR - http://www.scopus.com/inward/record.url?scp=0031778612&partnerID=8YFLogxK

U2 - 10.1099/00221287-144-6-1611

DO - 10.1099/00221287-144-6-1611

M3 - Article

AN - SCOPUS:0031778612

VL - 144

SP - 1611

EP - 1617

JO - Microbiology (United Kingdom)

JF - Microbiology (United Kingdom)

SN - 1350-0872

IS - 6

ER -