Marine methylotrophs revealed by stable-isotope probing, multiple displacement amplification and metagenomics.
Environ Microbiol
; 10(6): 1526-35, 2008 Jun.
Article
em En
| MEDLINE
| ID: mdl-18294205
ABSTRACT
The concentrations of one-carbon substrates that fuel methylotrophic microbial communities in the ocean are limited and the specialized guilds of bacteria that use these molecules may exist at low relative abundance. As a result, these organisms are difficult to identify and are often missed with existing cultivation and gene retrieval methods. Here, we demonstrate a novel proof of concept using environmentally-relevant substrate concentrations in stable-isotope probing (SIP) incubations to yield sufficient DNA for large-insert metagenomic analysis through multiple displacement amplification (MDA). A marine surface-water sample was labelled sufficiently by incubation with near in situ concentrations of methanol. Picogram quantities of labelled (13)C-DNA were purified from caesium chloride gradients, amplified with MDA to produce microgram amounts of high-molecular-weight DNA (
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Água do Mar
/
Bactérias
/
Técnicas Bacteriológicas
/
Técnicas de Amplificação de Ácido Nucleico
/
Genômica
Idioma:
En
Revista:
Environ Microbiol
Assunto da revista:
MICROBIOLOGIA
/
SAUDE AMBIENTAL
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Reino Unido