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Title of Journal: Arch Microbiol

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Abbravation: Archives of Microbiology

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Springer-Verlag

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DOI

10.1007/s001260050106

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ISSN

1432-072X

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Cloning and in vivo functional analysis by disrupt

Authors: KangMu Lee ChangKwon Lee SunUk Choi HaeRyong Park Shigeru Kitani Takuya Nihira YongIl Hwang
Publish Date: 2005/10/15
Volume: 184, Issue: 4, Pages: 249-257
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Abstract

A gene encoding a γbutyrolactone autoregulator receptor which has a common activity as DNAbinding transcriptional repressors controlling secondary metabolism and/or morphological differentiation in Streptomyces was cloned from a natamycin producer Streptomyces natalensis PCR using the primers designed for the two highly conserved regions of Streptomyces autoregulator receptors BarA FarA ScbR and ArpA gave a 102bp band The sequence of this band had a high similarity to the expected region of a receptor gene By genomic Southern hybridization with the 102bp insert as a probe a 687bp intact receptor gene sngR was obtained from S natalensis To clarify the in vivo function of sngR a sngRdisrupted strain was constructed and the phenotypes were compared with those of the wildtype strain The sngRdisruptants started natamycin production 6 h earlier and showed a 46fold higher production of natamycin than the wildtype strain In addition the sporulation began earlier and the number of spores was tenfold more abundant than that of the wildtype strain All the phenotypes were restored back to the original phenotypes of the wildtype strain by complementation with the intact sngR indicating that the autoregulator receptor protein of S natalensis acts as a primary negative regulator both on the biosynthesis of natamycin and sporulation

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