Leclercq Julie, Szabolcs Toth, Martin Florence, Montoro Pascal
CIRAD, UMR AGAP, F-34398 Montpellier, France.
Plasmid. 2015 Sep;81:50-4. doi: 10.1016/j.plasmid.2015.07.003. Epub 2015 Jul 23.
pCAMBIA vectors have become popular for their easy handling, stability and the existence of a range of selection and reporter genes. However, these vectors have yet to integrate the Gateway® cloning system, which has enabled site-specific recombination without the need for restriction enzymes and ligases. This paper sets out to convert the pCambia2300 binary vector into a destination vector with the Gateway® cassette driven by the CaMV35S promoter. The destination vector, pCamway35S, was then evaluated using the uidA reporter gene. Transient and stable transformation experiments were successfully assayed, either by particle bombardment or by Agrobacterium tumefaciens in Allium cepa and Hevea embryogenic calli. After counting the transformation units, the statistical analysis performed on the data showed that the pCamway 35S::uidA vector was as efficient as pCambia2301, a pCAMBIA2300 containing the uidA reporter gene under the CaMV 35S promoter.
pCAMBIA载体因其易于操作、稳定性以及一系列选择和报告基因的存在而受到欢迎。然而，这些载体尚未整合Gateway®克隆系统，该系统能够实现位点特异性重组，而无需限制性内切酶和连接酶。本文着手将pCambia2300双元载体转化为由CaMV35S启动子驱动的带有Gateway®盒的目的载体。然后使用uidA报告基因对目的载体pCamway35S进行评估。通过粒子轰击或根癌农杆菌在洋葱和橡胶树胚性愈伤组织中成功进行了瞬时和稳定转化实验。在对转化单位进行计数后，对数据进行的统计分析表明，pCamway 35S::uidA载体与pCambia2301一样高效，pCambia2301是一个在CaMV 35S启动子下含有uidA报告基因的pCAMBIA2300。
