| 根癌农杆菌介导里氏木霉转化体系的建立和条件优化 |
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| 引用本文： | 刘刚,杨长得,康康,邢苗,田生礼. 根癌农杆菌介导里氏木霉转化体系的建立和条件优化[J]. 生物技术, 2011, 21(4). DOI: 10.3969/j.issn.1004-311X.2011.04.098 |
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| 作者姓名： | 刘刚 杨长得 康康 邢苗 田生礼 |
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| 作者单位： | 深圳市微生物基因工程重点实验室,深圳大学生命科学学院,广东深圳518060) |
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| 基金项目： | 国家自然科学基金项目，深圳市科技计划项目 |
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| 摘 要： | 目的:采用根癌农杆菌介导的转化方法实现丝状真菌里氏木霉的遗传转化,并优化转化条件.方法:构建含潮霉素抗性基因(hph)的双元载体pCAM-hph后,转化根癌农杆菌LBA4404获得转化菌株.将根癌农杆菌的转化菌株和里氏木霉的分生孢子共培养后在含100μg/mL潮霉素的抗性平板上筛选里氏木霉转化子,并采用PCR扩增和序列测定对转化子中的插入片段进行了分析.结果:使用根癌农杆菌介导的转化方法转化里氏木霉,每106个分生孢子可获得25.8个转化子.最佳的转化条件为:农杆菌初始浓度为OD660约为0.8,孢子数为106个,共培养时间为48h,pH为5.0～5.5,培养温度为28℃.结论:建立了根癌农杆菌介导的里氏木霉转化方法,并获得了最佳的转化条件.
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| 关 键 词： | 里氏木霉 根癌农杆菌 分生孢子 潮霉素磷酸转移酶 双元载体 |
Agrobacterium tumefaciens Mediated Transformation of Trichoderma reesei and Optimization of the Transforming Conditions |
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| LIU Gang,YANG Chang-de,KANG Kang,XING Miao,TIAN Sheng-li. Agrobacterium tumefaciens Mediated Transformation of Trichoderma reesei and Optimization of the Transforming Conditions[J]. Biotechnology, 2011, 21(4). DOI: 10.3969/j.issn.1004-311X.2011.04.098 |
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| Authors: | LIU Gang YANG Chang-de KANG Kang XING Miao TIAN Sheng-li |
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| Affiliation: | LIU Gang,YANG Chang-de,KANG Kang,XING Miao,TIAN Sheng-li(Shenzhen Key Laboratory of Microbial Genetic Engineering,College of Life Sciences,Shenzhen University,Shenzhen 518060,China) |
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| Abstract: | Objective:Transformation of the filamentous fungus Trichoderma reesei was achieved using Agrobacterium tumeficiens-mediated transformation(ATMT) method,and the factors influencing transformation efficiency were evaluated.Method:The binary vector pCAM-hph containing the hygromycin phosphotransferase(hph) gene was constructed based on a commercial vector pCAMBIA 0380,and was transformed into A.tumeficiens LBA4404.The A.tumeficiens transformants were co-cultivated with the T.reesei conidial spores,and the fung... |
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| Keywords: | Trichoderma reesei Agrobacterium tumeficiens codinial spores hygromycin phosphotransferase binary vector |
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