期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 78, 期 2, 页码 150-154出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2009.05.005
关键词
Cosmid library; Gene screening; Organization strategy; Streptomyces; Tacrolimus
资金
- Ministry of Education and Science (Madrid, Spain) [BIO2006-14853-C02-01]
- Ministry of Education and Science
- Ministry of Science and innovation of Spain
- European Social Fund (ESF) [PTA-2003-01-00689]
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. (c) 2009 Elsevier B.V. All rights reserved.
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