Efficient pyramidal arrangement of an ordered cosmid library: Rapid screening of genes of the tacrolimus-producer Streptomyces sp. ATCC 55098.
J Microbiol Methods
; 78(2): 150-4, 2009 Aug.
Article
em En
| MEDLINE
| ID: mdl-19445977
ABSTRACT
A gene library is a useful tool for molecular biology studies, but the classical screening of multiple plates is laborious and time-consuming. Cosmid gene libraries are particularly well suited for isolation of large gene clusters encoding the biosynthetic pathways of secondary metabolites in Streptomyces. A gene library of the immunosuppressant tacrolimus-producer strain Streptomyces sp. ATCC 55098 was constructed in the SuperCos1 vector and 1656 clones were organized in an easy pyramidal arrangement system. This clustering method allows a dual efficient screening (PCR and in situ colony hybridization) of the gene library in a two-step method by using only one 96-well plate. The dual screening combines the advantages of both techniques, the swiftness of PCR and the robustness of colony hybridization.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Streptomyces
/
Cosmídeos
/
Biblioteca Gênica
/
Tacrolimo
/
Genes Bacterianos
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article