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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。
In order to cultivate transgenic wheat with high resistance and no selection marker, ThIPK2 gene was searched from NCBI database. Based on the amino acid sequence encoded by this gene, the gene was codon-optimized with reference to wheat preferred codon and repeated And unwanted restriction sites removed after artificial synthesis. The modified ThIPK2 gene was inserted between the strong promoter Ubiquitin and the terminator AtSac66 and then inserted into the plant expression vector containing the corn Ac / Ds transposon background skeleton. Maize Ubiquitin promoter drives plant expression vector for ThIPK2 gene. After restriction endonuclease analysis, the plant expression vector was successfully constructed.