Characterization of the dnaK Locus in Streptomyces griseus.
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概要
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The nucleotide sequence of the <I>dnaK</I> locus in <I>Streptomyces griseus</I> 2247 revealed a gene organization consisting of 6 genes (5'-<I>orf1</I>-<I>dnaK</I>-<I>grpE</I>-<I>dnaJ</I>-<I>hspR</I>-<I>orf6</I>-3'). The genes <I>orf1</I> and <I>orf6</I> were predicted to transcribe in the direction opposite that of <I>dnaK</I>, <I>grpE</I>, <I>dnaJ</I> and <I>hspR</I>. This locus of <I>S. griseus</I> was compared with that of <I>S. coelicolor</I> A3(2). No homolog of <I>orf1</I> and <I>orf6</I> was found in <I>S. coelicolor</I>, while <I>dnaK</I>, <I>grpE</I>, <I>dnaJ</I> <I>and</I> <I>hspR</I> were highly similar to those of <I>S. coelicolor</I>. Promoter probing analysis revealed significant promoter activity in the upstream region of the <I>dnaK</I> gene. The interspace regions of contiguous genes (<I>dnaK</I>-<I>grpE</I>-<I>dnaJ</I>-<I>hspR</I>) were amplified by RT-PCR of mRNA, indicating that these four genes constitute a polycistronic operon. His-tagged HspR protein was overexpressed in <I>Escherichia coli</I> and purified.
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