EPSPS基因克隆及其表达载体的构建

【目的】克隆大豆EPSPS基因并构建其表达载体,为培育抗草甘膦作物提供理论基础和技术储备。【方法】以抗草甘膦大豆总基因组为模板,扩增EPSPS基因,构建EPSPS基因的植物表达载体PCAMBIA1300-UBI-GFP-EPSPS,并导入农杆菌,使其能够进行作物的遗传转化。【结果】扩增获得EPSPS基因全长1368bp,共编码455个氨基酸。测序结果表明,其与GenBank（AF464188.1）中已知的CDS序列完全一致,成功构建了EPSPS植物表达载体,并将其导入农杆菌菌株EHA105中。【结论】构建的表达载体可用于甜高粱等单子叶作物抗草甘膦性状的遗传改良。...

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Bibliographic Details
Published in南方农业学报 Vol. 43; no. 9; pp. 1257 - 1261
Main Author 谢鸿锴 禤维言 王磊 冯斗
Format Journal Article
LanguageChinese
Published 广西大学农学院,南宁,530005 2012
Subjects
EPSPS基因
大豆
抗草甘膦
构建
表达载体
soybean
EPSPS基因
表达载体
大豆
construction
构建
expression vector
抗草甘膦
EPSPS gene
glyphosate resistance
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ISSN2095-1191
DOI10.3969/j:issn.2095-1191.2012.09.1257

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Summary:【目的】克隆大豆EPSPS基因并构建其表达载体,为培育抗草甘膦作物提供理论基础和技术储备。【方法】以抗草甘膦大豆总基因组为模板,扩增EPSPS基因,构建EPSPS基因的植物表达载体PCAMBIA1300-UBI-GFP-EPSPS,并导入农杆菌,使其能够进行作物的遗传转化。【结果】扩增获得EPSPS基因全长1368bp,共编码455个氨基酸。测序结果表明,其与GenBank（AF464188.1）中已知的CDS序列完全一致,成功构建了EPSPS植物表达载体,并将其导入农杆菌菌株EHA105中。【结论】构建的表达载体可用于甜高粱等单子叶作物抗草甘膦性状的遗传改良。
Bibliography:XIE Hong-kai,XUAN Wei-yan,WANG Lei,FENG Dou（Agricultural College,Guangxi University,Nanning 530005,China）
45-1381/S
[Objective]This research aimed to clone EPSPS gene of soybean and construct its expression vector in order to provide the theoretical basis and the technical reservation for the cultivation of glyphosate-resistant crop.[Method]The total genome of glyphosate-resistant soybean was used as templates.The EPSPS gene was amplified,and its plant expression vector,PCAMBIA1300-UBI-GFP-EPSPS,was constructed;afterwards,it was translated into Agrobacterium tumefaciens in order to carry out the crop＇s genetic transformation.[Result]The full-length of the amplified EPSPS gene was 1368 bp,which encoded 455 amino acids.The sequencing result showed the identical CDS sequence as the known one in GenBank（AF464188.1）.The plant expression vector of EPSPS was constructed successfully and translated into Agrobacterium tumefaciens EHA105.[Conclusion]After its genetic transformation,this expression vector could be used fo
ISSN:2095-1191
DOI:10.3969/j:issn.2095-1191.2012.09.1257