玉米C4-PEPC基因RNAi植物表达载体的构建

Construction of Plant Transformation Vector of Maize C4-PEPC Gene for RNAi

玉米C4光合关键酶PEPC基因在玉米C4光合作用中起到重要作用,根据已测序的玉米C4-PEPC基因序列,设计2对带限制性内切酶位点的特异性引物,以pBI121 C4-PEPC质粒DNA为模板,PCR扩增得到2个C4-PEPC基因片段。再将正向和反向两个片段分别经双酶切后插入植物表达载体pTA7002的XhoI和SpeI酶切位点之间,从而构建了以玉米C4-PEPC基因为靶标的RNAi植物表达载体p7002-Z-F。载体质粒经酶切鉴定正确后通过冻融法转入根癌农杆菌EHA105中,为后续探讨干扰载体的干涉PEPC基因表达效果奠定了基础。

Maize C4-PEPC gene plays an important role in the C4 photosynthesis of it,Two pairs of primers containing restriction enzyme site were designed and used to amplify sequences from pBI121 C4-PEPC,. Two PCR products were double digested by corresponding enzymes respectively and inserted between XhoI and SpeI enzyme sites of expression vector pTA7002. Confirmed by restriction endonucleases, the RNAi expression vector was transformed into Agrobacterium tumefaciens strain EHA105 by freezing-thaw method, it provided an important basis for further studying its RNAi effeetion.