期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 100, 期 6, 页码 2797-2808出版社
SPRINGER
DOI: 10.1007/s00253-015-7271-0
关键词
Streptomyces; Phi FBT1 integrative vector; Heterologous expression; Neutral phenotype
资金
- ERC [280304]
- Severo Ochoa fellowship (FICYT, Consejeria de Educacion y Ciencia, Spain)
- European Research Council (ERC) [280304] Funding Source: European Research Council (ERC)
Integrative plasmids are one of the best options to introduce genes in low copy and in a stable form into bacteria. The I broken vertical bar C31-derived plasmids constitute the most common integrative vectors used in Streptomyces. They integrate at different positions (attB and pseudo-attB sites) generating different mutations. The less common I broken vertical bar BT1-derived vectors integrate at the unique attB site localized in the SCO4848 gene (S. coelicolor genome) or their orthologues in other streptomycetes. This work demonstrates that disruption of SCO4848 generates a delay in spore germination. SCO4848 is co-transcribed with SCO4849, and the spore germination phenotype is complemented by SCO4849. Plasmids pNG1-4 were created by modifying the I broken vertical bar BT1 integrative vector pMS82 by introducing a copy of SCO4849 under the control of the promoter region of SCO4848. pNG2 and pNG4 also included a copy of the P (ermE) * in order to facilitate gene overexpression. pNG3 and pNG4 harboured a copy of the bla gene (ampicillin resistance) to facilitate selection in E. coli. pNG1-4 are the only integrative vectors designed to produce a neutral phenotype when they are integrated into the Streptomyces genome. The experimental approach developed in this work can be applied to create phenotypically neutral integrative plasmids in other bacteria.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据