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【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。
【Objective】 The EPSPS gene of soybean was cloned and its expression vector was constructed, which provided the theoretical basis and technical reserve for the cultivation of glyphosate resistant crop. 【Method】 The EPSPS gene was amplified from glyphosate-tolerant soybean genome and the plant expression vector PCAMBIA1300-UBI-GFP-EPSPS of EPSPS gene was constructed and introduced into Agrobacterium tumefaciens so that it could be used for genetic transformation of crops. 【Result】 The EPSPS gene was amplified to 1368bp in length and encoded a total of 455 amino acids. Sequencing results showed that it was completely identical with the known CDS sequence in GenBank (AF464188.1). EPSPS plant expression vector was successfully constructed and introduced into Agrobacterium tumefaciens strain EHA105. 【Conclusion】 The constructed expression vector can be used for genetic improvement of glyphosate-resistant traits in monocotyledonous plants such as sweet sorghum.