Efficient pyramidal arrangement of an ordered cosmid library: Rapid screening of genes of the tacrolimus-producer Streptomyces sp. ATCC 55098.
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.
Martínez-Castro
Miriam
M
Microbiology Area, Molecular Biology Department, Biology and Environmental Sciences Faculty, Campus de Vegazana s/n, University of León, 24071 León, Spain.
Solera
Elena
E
Martín
Juan F
JF
Barreiro
Carlos
C
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*
D004269:DNA, Bacterial; D016559:Tacrolimus