机构地区: 仲恺农业工程学院生命科学学院
出 处: 《中国农学通报》 2009年第12期25-29,共5页
摘 要: 为了解细极链格孢菌(Alternaria tenuissima)遗传学背景,克隆及研究该菌功能基因,根据Gateway技术构建了既能在酵母细胞中表达外源基因又能在原核细胞大量复制的酵母表达载体pRS-DEST42,构建了细极链格孢菌(A.tenuissima)cDNA酵母表达文库。经检测,文库的平均滴度为2.44×106cfu/ml,文库总容量为2.44×107cfu(colony-forming unit),阳性克隆率为100%,平均插入片段约为1.38kb。细极链格孢菌cDNA酵母表达文库是一个质量较高的表达文库,为克隆与分离全长目的基因及其功能奠定了坚实的基础。 For further understand genetics background and functional genes of A. tenuissima. A yeast destination vector pRS-DEST42, express foreign genes in a low-copy number in yeast and replicate high-copy in E. coli, was constructed. The cDNA yeast expression library of A. tenuissima was constructed through Gateway system. It has a higher titer of 2.44×10^6 cfu/ml and a total clones of 2.44×10^7 cfu. The rate of positive recombinants clones was 100% and the size of average insert cDNA was 1.38 kb. This provides a basis for isolate and study on functional genes in A. tenuissima.