Overexpression of regulatory genes in S. nodosus. a The map of plasmid vector pTMW50 used for overexpression. This plasmid contains a constitutively expressed promoter ermE. The restriction sites BamHI and XbaI were used for cloning. b Plasmid construction strategy used in the overexpression of regulatory genes
Plasmid pTMW50, an integrative vector that harbors the
constitutive expression promoter ermE (Fig. 5a), was used as
the overexpression vector for the regulatory gene(s) in S.
The asu1–asu4 gene locus, located upstream of the asukamycin
biosynthetic gene cluster, was cloned into the integration vector
pTMW50 (Fig. 5a), a pSET152 derivative containing the aac3(IV)
gene and an ermE promoter, to give pTMPX75 (Fig. 5b).
The genes were individually cloned into the vector pTMW50 to
generate plasmids pTMPX82, pTMPX83, pTMPX84, and pTMPX85
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