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质粒pIJ4026和pRK404-vhb分别含有红霉素抗性基因启动子(PermE)和透明颤菌血红蛋白基因(vhb),先用PCR分别扩增二者,再用SOE-PCR(Splicing by overlap extension PCR)将不含自身启动子的vhb基因置于PermE之下;将SOE-PCR产物克隆到链霉菌整合性载体pSET152上,构建成vhb基因表达载体pWQ2004.通过大肠埃希氏菌-链霉菌属间接合转移的方式将pWQ2004导入弗氏链霉菌(Streptomyces fradiae)中,利用pWQ20