摘要
采用基于EST的电子克隆方法,以拟南芥(Arabidopsis thaliana)的尿黑酸叶绿基转移酶基因(Homogentisate phytyltransferase,HPT)cDNA序列为信息探针,对大豆的EST数据库进行同源检索筛选,获得了1 777 bp长大豆HPT基因的cDNA序列(GenBank登录号为:AY956421),经RT-PCR扩增、分子克隆和序列分析验证,表明与电子克隆序列一致;该基因具有完整的开放阅读框架(ORF,173～1 408 bp),推测编码411个氨基酸的蛋白。该蛋白与拟南芥(Ara-bidopsis thaliana)、苜蓿(Medicago sativa)、水稻(Oryza sativa)、玉米(Zea mays)、光合集胞蓝细菌(Synechocystis)的序列进行了比较,发现该基因具有保守性。
In silico cloning based on EST sequence is a new method of gene cloning, using Arabidopsis thaliana homogentisate phytyltransferase cDNA sequence as a query probe to blast Glycine max EST database, a soybean HPT gene cDNA of 1 777 bp was obtained（GeneBank Accession, AY956421）. This sequence was confirmed by RT-PCR, molecular cloning and sequencing, It contained a complete ORF, from 173 bp to 1 408 bp, encoded 411 amino acids, and conserved with Arabidopsis thaliana , Medicago sative , Oryza , Zea mays and Synechocystis . The results show that it is an efficient technique to clone new genes by searching EST database with homologous gene of model species.
出处
《华北农学报》
CSCD
北大核心
2007年第4期14-18,共5页
Acta Agriculturae Boreali-Sinica
基金
北京市教委科技发展计划项目基金资助(200310028112)
