论文部分内容阅读
牻牛儿基牻牛儿基焦磷酸合成酶是红豆杉细胞紫杉醇生物合成途径中的关键酶.试验采用RT-PCR技术从东北红豆杉中克隆GGPPS基因片段,将GGPPS基因片段与pMD20-Tvector连接,转化E.coli DH5α,对阳性克隆进行序列测定.生物信息学分析结果表明,获得GGPPS基因片段长度为371 bp,与GenBank中收录的GGPPS同源性达到99%.为从内生真菌紫杉醇生物合成途径中克隆关键酶基因和高产紫杉醇基因工程菌株的构建提供了借鉴.“,”Geranylgeranyl diphosphate synthase(GGPPS) is a key enzyme in the taxol biosynthesis.Fragment of GGPPS gene was obtained from Taxus cuspidata by RT-PCR,and it was cloned in pMD20-T vector,then it was transformed into E.coli DH5α.A 371 bp fragment of GGPPS gene was obtained by sequencing after screening the positive clones by PCR.Its sequence had 99% similarity with GGPPS embodied in GenBank.The study laid solid foundation for cloning of genes related to taxol biosynthesis and construction of engineering strains with high yield of taxol by genetic techniques.