vgb在红色糖多孢菌表达及生物活性的研究  

Study on Expression and Activity of Vgb in Recombinant Sac. Erythraea

作　　者：吴益民[1] 王洪军[1] 孙艳[1] 王立强[1] 张志强[1] 冯立[1] 杨青[1] 

机构地区：[1]沈阳军区疾病预防控制中心,辽宁沈阳110034

出　　处：《药物生物技术》2007年第2期90-93,共4页Pharmaceutical Biotechnology

基　　金：辽宁省教育厅青年人才基金资助课题(No.05L433)

摘　　要：利用基因工程技术对红色糖多孢菌进行改造,提高红霉素产率。用PCR技术克隆vgb,采用电穿孔法与红色糖多孢发酵菌染色体整合,鉴定采用Western blot与Southern blotting分析。VHb活性分析采用一氧化碳(CO)差示光谱法,红霉素效价测定采用管碟法。结果克隆了含vgb的红色糖多孢菌表达质粒(pBlueV),筛选了重组红色糖多孢菌株。与原始菌株比较,重组菌株细胞发酵密度分别为1.37与2.82,红霉素效价分别为3.8与5.1,相当于重组菌株提高红霉素体积产率约29%。重组红色糖多孢发酵菌提高了红霉素产率,对解决抗生素工业和基因工程菌高密度发酵有良好的应用前景。In order to promote the production of erythromycin and to reduce cost in manufacture, Sac. Erythraea was improved by gene recombinant technology, vgb was cloned by PCR, and vgb was integrated into the chromosomal DNA of Sac. erythraea by electrotransformation. The VHb was verified by analysis of Western blotting and Southern blotting. Activity of VHb was examined by CO different spectra. The cell density and erythromycin titer were determined by sepetrodensitometer and pipe-dish respectively. The results showed that the cell density and Erythromycin titer in riginal strain and in recombicant strain were 1.37, 3.8 and 2. 82, 5.1 respectively. Recombinant Sac. Erythraea has raised erythromycin output,it will benefit for the antibiotics industry production in high-cell density fermentation.

关 键 词：血红蛋白基因 红色糖多孢菌 红霉素 高密度发酵 

分 类 号：Q753[生物学—分子生物学]