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作 者:薛姣[1] 胡宗利[1] 陈国平[1] 赵志平[1] 陈绪清[1]
出 处:《广东农业科学》2008年第6期83-87,共5页Guangdong Agricultural Sciences
基 金:国家"863"计划资助项目(2006AA02Z138);重庆市科委自然科学基金重点项目(CSTC;2006BA5006)
摘 要:研究利用PCR技术扩增出Erwinia herbicola的crtI基因,并连接到含强启动子Puc的表达载体pRKR5上构建表达质粒pRKR5-crtI,通过接合转移的方式将其导入光合细菌Rhodobacter sphaeroides突变株TC72中,调控氧浓度诱导工程菌累积红色色素,经HPLC和吸收光谱分析,工程菌中合成的色素为番茄红素,工程菌的生物量(干重)为2.36 g/L,番茄红素含量可达1.52 mg/g,与其他生产番茄红素的工程菌如大肠杆菌和产朊假丝酵母相比,番茄红素产量有一定程度的提高。In this research, the crtl of Erwinia herbicola was amplified by polymerase chain reaction and cloned into pRKR5 vector controlled by puc promoter from Rhodobacter sphaeroides, to construct pRKRS-crtI expression cassette. Then the expression construct was introduced into photosynthetic mutant bacteria TC72 by conjugative transfer. Induced by semiaerobic and aerobica conditions, the transformants could express crtl high efficiently and accumulate a red pigment quickly. The red pigment was testified as lycopene by absorbance spectra analysis and HPLC analysis. The engineering bacteria could grow up to 2.36 g/L and accumulate lycopene to 1.52 mg/g. Compared with other engineering bacteria producing lycopene, such as Escherichia coli and Candida utilis, the output of lycopene in the transformant had some increase.
关 键 词:番茄红素 crtI基因 ERWINIA herbicola RHODOBACTER SPHAEROIDES 异源表达
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