摘要
采用RT-PCR技术从乌拉尔甘草(Glycyrrhiza uralensis)中克隆甘草鲨烯合成酶(Squalene Syn-thase,SS)基因,并通过酶切连接的方法构建相关植物表达载体。结果表明,两个SS基因编码区长分别为1242bp、1239bp,分别编码412、413个氨基酸残基的多肽,与Hayashi等报道的光果甘草两个SS基因(GgSQS1和GgSQS2)同源性高达98%,分别命名为GuSQS1和GuSQS2(GenBank登录号分别为:AM182329,AM182330);植物表达载体构建的鉴定结果表明,已将GuSQS1和GuSQS2序列分别正向插入到双T-DNA表达载体中的CaMV 35S启动子和NOS终止子之间,重组质粒分别命名为130/35S-GuSQS1和130/35S-GuSQS2,为今后的次生代谢基因工程研究奠定基础。
Two squalene synthase（SS）genes were cloned from Glycyrrhiza uralensis using RT-PCR method and constructed their expression vectors by restriction endonucleases and ligases. The results showed that two GuSQS were 1142bp and 1139bp in length encoding two pelypeptides of 413 and 412 amino acids respectively sharing 98% identity to two SS cDNA from Glycyrrhiza glabra reported by Hayashi and were named as GgSQS1 and GgSQS2 （GenBank Accession number. AM182329, AM182330）, respectively. Analysis of constructions verified that GgSQS1 and GgSQS2 were inserted between CaMV35S promoter and NOS terminator in double T-DNA expression vector. Two recombinants gained were designated as 130/35S-GuSQS1 and 130/35S-GuSQS2, respectively, which would lay a foundation for studying gene engineering of secondary metabolism in future.
出处
《药物生物技术》
CAS
CSCD
2007年第4期255-258,共4页
Pharmaceutical Biotechnology
关键词
乌拉尔甘草
甘草酸
鲨烯合成酶
基因克隆
双T-DNA表达载体
Glycyrrhiza uralensis, Glycyrrhizic acid, Squalene synthase Gene cloning, Double T-DNA expression vector
