ThIPK2基因植物表达载体的构建

为培育高度抗逆和无选择标记的转基因小麦,本研究从NCBI数据库中搜索到ThIPK2基因序列,依据该基因编码的氨基酸序列,参照小麦偏爱的密码子对该基因进行密码子优化,并将重复的和不必要的酶切位点去掉后进行人工合成。将改造后的ThIPK2基因插入到强启动子Ubiquitin和终止子AtSac66之间,然后将其插入到含有玉米Ac/Ds转座子背景骨架的植物表达载体中,获得了具有删除选择标记功能的、由玉米Ubiquitin启动子驱动ThIPK2基因的植物表达载体。经过限制性内切酶分析鉴定,该植物表达载体构建成功。

Construction of Plant Expression Vector Harboring ThIPK2

In order to get stress tolerant transgenic wheat plants without selective markers,the ThIPK2 gene sequence was got from the NCBI database,the gene codons were optimized based on wheat preference codons,the repeated and unnecessary restriction sites were removed,and the gene was synthesized. The synthesized ThIPK2 gene was constructed into vector with Ubiquitin promoter and AtSac66 terminator. The whole cassete was then ligated into a plant expression vector harboring Ac/Ds transposon,which could be further used for wheat transformation. Identified by restriction endonuclease,the expression vector was successfully constructed.