银杏EPSPS基因克隆及表达分析  

Cloning and Expression Analysis of EPSP Synthase Gene from Ginkgo biloba L.

作　　者：程华[1,2] 李琳玲[2] 王燕[3] 姜德志[3] 程水源[1,2] 

机构地区：[1]南京林业大学森林资源与环境学院,南京210037 [2]黄冈师范学院生命科学与工程学院,湖北黄冈438000 [3]长江大学园艺园林学院,湖北荆州434025

出　　处：《西北植物学报》2010年第12期2365-2372,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(30971974);湖北省自然科学基金计划重点项目(2008CDA061)

摘　　要：利用RACE技术,克隆到银杏EPSPS合酶基因(GbEPSPS)的cDNA序列并对其进行生物信息学分析。结果表明:银杏GbEPSPScDNA长1 403 bp(GenBank登录号GU084139),其包含一个1 035 bp的ORF框,编码344个氨基酸;预测该基因编码蛋白质分子量为36.87 kD,其等电点为5.75。系统进化树分析表明,银杏EPSPS蛋白质序列与其他物种的EPSPS同源性较高。半定量RT-PCR分析结果显示,EPSPS基因在银杏的叶和果实中表达量最高,其次为茎,根中表达水平最低。草甘膦处理能显著诱导银杏EPSPS基因表达量升高;紫外光对银杏EPSPS基因的表达具有诱导作用;ABA诱导GbEPSPS表达量先升后降;GbEPSPS转录水平受到42℃高温显著诱导。The RACE technology was used for cloning the full-length cDNA of EPSP synthase gene from Ginkgo biloba.The cDNA sequence was 1 403 bp with a poly A tail,and contained a 1 035 bp open reading frame（ORF） encoding a 344 amino acid protein.Bioinformatics analysis predicted that it coded a protein of 36.87 kD,and its isoelectric point was 5.75.Phylogenetic tree analysis showed that the Ginkgo biloba EPSP synthase protein sequences was higly homology with the EPSP synthase of other species.RT-PCR analysis showed that GbEPSPS expressed in leaves,stems,roots and fruits,and had the highest expression in leaves and fruits,the next in stems.The least expression had been found in roots.The expression of GbEPSPS could be induced by glyphorose and UV-B.ABA could improve the expression of GbEPSPS first,but deduce later.The transcription levels were significantly induced by 42℃.

关 键 词：银杏 EPSP合酶 基因克隆 RACE技术 

分 类 号：Q789[生物学—分子生物学]