| 大豆尿黑酸叶绿基转移酶基因的克隆与进化分析 |
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| 引用本文： | 孟宪萍,李晓晓,李雅轩,蔡民华,胡英考.大豆尿黑酸叶绿基转移酶基因的克隆与进化分析[J].华北农学报,2007,22(4):14-18. |
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| 作者姓名： | 孟宪萍 李晓晓 李雅轩 蔡民华 胡英考 |
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| 作者单位： | 首都师范大学,生命科学学院,北京,100037 |
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| 基金项目： | 北京市教委科技发展计划项目基金资助(200310028112) |
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| 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。
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| 关 键 词： | 电子克隆 EST数据库 RT-PCR 尿黑酸叶绿基转移酶 大豆 |
| 文章编号： | 1000-7091（2007）04-0014-05 |
| 修稿时间： | 2007-04-16 |
In silico Cloning of Homogentisate Phytyltransferase Gene from Soybean |
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| MENG Xian-ping,LI Xiao-xiao,LI Ya-xuan,CAI Min-hua,HU Ying-kao.In silico Cloning of Homogentisate Phytyltransferase Gene from Soybean[J].Acta Agriculturae Boreali-Sinica,2007,22(4):14-18. |
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| Authors: | MENG Xian-ping LI Xiao-xiao LI Ya-xuan CAI Min-hua HU Ying-kao |
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| Institution: | College of Life Science, Capital Normal University, Beijing 100037, China |
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| Abstract: | 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. |
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| Keywords: | In silico cloning EST database RT-PCR Homogentisate phytyltransferase Soybean |
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