里氏木霉外源表达载体的构建

摘要

[ Objecfve ] The study was to construct a Trichoderma reesei vector for expressing external genes. [ 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 6 strains capable of growing on basic medium containing 175 mg/L hygromycin B, further conducted hygromycin resistance test. [ Result] In comparison with the original strain( wild type), hygromycin resistance the 6 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.%[目的]构建里氏木霉外源表达栽体.[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外源表达栽体,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175 mg/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,证明表达栽体构建成功.[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.