Wide host-range cloning for functional metagenomics.
Methods Mol Biol
; 668: 77-96, 2010.
Article
em En
| MEDLINE
| ID: mdl-20830557
ABSTRACT
We describe how wide host-range cloning vectors can lead to more flexible and effective procedures to isolate novel genes by screening metagenomic libraries in a range of bacterial hosts, not just the conventionally used Escherichia coli. We give examples of various wide host-range plasmid, cosmid, and BAC cloning vectors and the types of genes and activities that have been successfully obtained to date. We present a detailed protocol that involves the construction and screening of a metagenomic library comprising fragments of bacterial DNA, obtained from a wastewater treatment plant and cloned in a wide host-range cosmid. We also consider future prospects and how techniques and tools can be improved.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Clonagem Molecular
/
Metagenoma
/
Metagenômica
/
Vetores Genéticos
Tipo de estudo:
Guideline
Idioma:
En
Revista:
Methods Mol Biol
Ano de publicação:
2010
Tipo de documento:
Article