Construction of Escherichia coli-Streptomyces shuttle expression vectors for gene expression in Streptomyces

pIJ4123 and pIJ6021 are high-copy-number vectors for gene expression in Streptomyces. They contain a strong, thiostrepton-inducible promoter, PtipA. Two E. coli-Streptomyces shuttle vectors, pHZ1271 and pHZ1272, were constructed by inserting a replicon and bla gene from E. coli into pIJ4123 and pIJ6021, respectively. Both vectors were structurally stable either in E. coli or in Streptomyces lividans. To demonstrate the utility of pHZ1272, the hemoglobin gene of Vitreoscillia (vhb) was cloned into pHZ1272 and expressed in Streptomyces lividans. The expression product (VHb) was detected by Western blotting and carbon monoxide binding activity analysis.