摘要
[目的]构建里氏木霉外源表达载体。[方法]以里氏木霉40359的CBHI启动子和终止子构建了里氏木霉40359外源表达载体,并用该表达载体成功表达了抗潮霉素B磷酸转移酶基因,得到6株可在含175mg/L潮霉素的基础培养基上生长的抗性菌株;提取6株抗性菌株的DNA整合到里氏木霉菌株中,并对其进行潮霉素耐受性检测。[结果]6株抗性菌株的抗潮霉素能力比原始菌株提高了75%;转基因菌株的潮霉素抗性可以稳定遗传,证明表达载体构建成功。[结论]为开展里氏木霉分子生物学研究与遗传改造奠定了基础。
[Objective] The study was to construct a exogenous expression vector for Trichoderma reesei.[Method] Using CBHI promoter and terminator of T.reesei strain 40359,we constructed an expression vector of T.reesei strain 40359 for expressing Hpt gene and got six strains capable of growing on basic medium containing 175 mg/L of hygromycin B,further conducted hygromycin resistance test.[Results] In comparison with the original strain（wild type）,hygromycin resistance the six engineered strains was increased by 75%;the hygromycin resistance could inherit stably.[Conclusion] Our results laid basis for biological study on T.reesei at molecular and genetically engineering levels.
关键词
里氏木霉
载体
遗传转化
Trichoderma reesei
Vector
Genetic transformation
