水稻细菌性条斑病非寄主抗性基因Rxo1的双右边界双元载体的构建

Construction of Double Right-Border Binary Vector Carrying Non-Host Gene Rxo1 Resistant to Rice Bacterial Leaf Streak

Rxo1是从玉米中克隆的水稻细菌性条斑病非寄主抗性基因。采用ScaⅠ和NgoMⅣ酶切携带Rxo1基因的载体pCAMBIA1305-1,HpaⅠ和XmaⅠ酶切双右边界双元载体pMNDRBBin6,通过平端和黏性末端连接目的基因和载体,得到了重组载体pMNDRBBin6-Rxo1。PCR和限制性内切酶酶切以及序列测定证实Rxo1整合到了双右边界载体pMNDRBBin6中。该载体为利用转基因技术获得无选择标记抗细菌性条斑病水稻育种材料提供了条件。

Rxol cloned from maize was a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxol was digested with Sca Ⅰ and NgoM IV and the double right border binary vector pMNDRBBin6 was digested with Hpa Ⅰ and Xma Ⅰ. pMNDRBBin6 carrying the gene Rxol was acquired by ligation of blunt-end and cohesive end. The resuits of PCR, restriction digestion and sequencing showed that the Rxol gene has been cloned into pMNDRBBin6. This double right-border binary vector, being named as pMNDRBBin6-Rxol , would play a role in breeding marker-free plants resistant to rice bacterial leaf streak by genetic transformation.