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KMID : 0545120090190020121
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Journal of Microbiology and Biotechnology
2009 Volume.19 No. 2 p.121 ~ p.127
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Identification and Functional Characterization of an afsR Homolog Regulatory Gene from Streptomyces venezuelae ATCC 15439
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Maharjan Sushila
Oh Tae-Jin
Lee Hei-Chan
Sohng Jae-Kyung
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Abstract
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Sequencing analysis of a 5-kb DNA fragment from Streptomyces venezuelae ATCC 15439 revealed the presence of one 3.1-kb open reading frame (ORF), designated afsRsv. The deduced product of afsR-sv (1,056 aa) was found to have high homology with the global regulatory protein AfsR. Homology-based analysis showed that afsR-sv represents a transcriptional activator belonging to the Streptomyces antibiotic regulatory protein (SARP) family that includes an Nterminal SARP domain containing a bacterial transcriptional activation domain (BTAD), an NB-ARC domain, and a Cterminal tetratricopeptide repeat domain. Gene expression analysis by reverse transcriptase PCR (RT-PCR) demonstrated the activation of transcription of genes belonging to pikromycin production, when afsR-sv was overexpressed in S. venezuelae. Heterologous expression of the afsR-sv in different Streptomyces strains resulted in increased production of the respective antibiotics, suggesting that afsR-sv is a positive regulator of antibiotics biosynthesis.
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KEYWORD
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afsR-sv, regulatory gene, reverse transcriptase PCR, SARP family, Streptomyces venezuelae
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