title: Efficient pyramidal arrangement of an ordered cosmid library: Rapid screening of genes of the tacrolimus-producer Streptomyces sp. ATCC 55098.
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.
authors.affiliation: Microbiology Area, Molecular Biology Department, Biology and Environmental Sciences Faculty, Campus de Vegazana s/n, University of León, 24071 León, Spain.
authors.forename: Miriam
authors.initials: M
authors.lastname: Martínez-Castro
authors.forename: Elena
authors.initials: E
authors.lastname: Solera
authors.forename: Juan F
authors.initials: JF
authors.lastname: Martín
authors.forename: Carlos
authors.initials: C
authors.lastname: Barreiro
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D003360:Cosmids; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D016559:Tacrolimus / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D016559:Tacrolimus