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Applied and Environmental Microbiology, March 2003, p. 1647-1654, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1647-1654.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of the Streptomyces sp. Strain C5 snp Locus and Development of snp-Derived Expression Vectors

Charles L. DeSanti1* and William R. Strohl2,{dagger}

Department of Microbiology, The Ohio State University, Columbus, Ohio 43210,1 Natural Products Microbiology, Merck Research Laboratories, Rahway, New Jersey 070652

Received 8 May 2002/ Accepted 15 December 2002

The Streptomyces sp. strain C5 snp locus is comprised of two divergently oriented genes: snpA, a metalloproteinase gene, and snpR, which encodes a LysR-like activator of snpA transcription. The transcriptional start point of snpR is immediately downstream of a strong T-N11-A inverted repeat motif likely to be the SnpR binding site, while the snpA transcriptional start site overlaps the ATG start codon, generating a leaderless snpA transcript. By using the aphII reporter gene of pIJ486 as a reporter, the plasmid-borne snpR-activated snpA promoter was ca. 60-fold more active than either the nonactivated snpA promoter or the melC1 promoter of pIJ702. The snpR-activated snpA promoter produced reporter protein levels comparable to those of the up-mutated ermE* promoter. The SnpR-activated snpA promoter was built into a set of transcriptional and translational fusion expression vectors which have been used for the intracellular expression of numerous daunomycin biosynthesis pathway genes from Streptomyces sp. strain C5 as well as the expression and secretion of soluble recombinant human endostatin.

* Corresponding author. Present address: Battelle Memorial Institute, 505 King Ave., Columbus, OH 43201-2693. Phone: (614) 424-5534. Fax: (614) 424-4185. E-mail: desantic{at}battelle.org.

{dagger} Present address: Department of Biologics Research, Merck Research Laboratories, Rahway, NJ 07065.

Applied and Environmental Microbiology, March 2003, p. 1647-1654, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1647-1654.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Chen, Y., Wendt-Pienkowski, E., Shen, B. (2008). Identification and Utility of FdmR1 as a Streptomyces Antibiotic Regulatory Protein Activator for Fredericamycin Production in Streptomyces griseus ATCC 49344 and Heterologous Hosts. J. Bacteriol. 190: 5587-5596 [Abstract] [Full Text]  


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