大肠杆菌链霉菌假单胞菌穿梭表达BAC载体的构建

Construction of an E.coli-Streptomyces-Pseudomonas Shuttle Expression BAC Vector

异源表达生理活性物质的生物合成基因簇是药物研发领域的一个重要的方向,它可通过异源表达来研究基因功能,获得目标化合物或对现有的化合物进行结构改造.生物合成基因簇一般较大,常规的DNA载体由于容量不足、拷贝数较低或不能在不同的异源宿主间穿梭表达而难以对其进行操作.本研究运用重组工程策略和常规的克隆手段,将多个异源表达所必须的功能基因克隆至常用的构建BAC文库的载体pECBAC1,获得了一个可在大肠杆菌链霉菌假单胞菌3个常见的异源表达宿主间进行穿梭表达的BAC载体.

Heterologous expression of biosynthetic gene cluster is an important study area of drug research and development,and it can be used for gene function study,to obtain target compound or its derivatives.As biosynthetic gene cluster is usually very large,normal DNA vectors may have the problems of low content,hard to handle because of vector＇s low copy number or not suit for shuttle expression in different heterologous strains.In this work,recombineering strategy as well as normal gene cloning techniques are used to clone functional heterologous expression DNA elements into pECBAC1,one of the most commonly used BAC library construction vector.