基于In-fusion技术的ERECTA基因植物表达载体的构建

Construction of ERECTA Gene Plant Expression Vector that Based on In-fusion Technology

以黄瓜品种‘长春密刺’(CCMC)为试材,利用高保真酶Iproof及不依赖于酶切的In-fusion技术,构建基于本底启动子驱动的黄瓜ERECTA基因的植物表达载体,以探讨ERECTA基因在黄瓜中的功能,以期为黄瓜抗性性状的遗传改良提供技术支持。结果表明:从CCMC的基因组DNA中克隆的2段gERECTA(DNA序列)长度分别为8 940bp和9 812bp,并分别命名为CsgERECTA-Flag和CsgERECTA-Poly。经过质粒PCR、梯度片段PCR和测序结果的鉴定表明,pCAMBIA3301-gERECTA的2个植物表达载体都已成功构建并转入根癌农杆菌EHA105中。

Taking cucumber breed of‘Changchun Mici＇（CCMC）as material,using high fidelity polymerase,Iproof and In-fusion technology that not based on digestion to construct plant expression vector of ERECTAgene that was driven by its own promoter for discussing the function of ERECTAgene and providing technology to improve resistance inheritance in cucumber.The results showed that two segments amplified from‘CCMC＇genome DNA were named CsgERECTA？？Flag which was 8 940 bp and CsgERECTA-Poly which was 9 812 bp.Plasmid PCR,gradient segments PCR and sequencing were conducted,which showed that two pCambia3301-gERECTA plant expression vector were both constructed and transformed into Agrobacterium tumefaciens EHA105.