大豆蛋白酶抑制剂基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Soybean Proteinase Inhibitor Gene

为了研究使用安全的抗虫蛋白,以市售吉育47种子为材料,利用RT-PCR方法,从大豆吉育47萌发种子提取RNA,以该RNA为模板进行反转录并进行PCR,扩增大豆吉育47蛋白酶抑制剂基因并进行生物信息学分析与表达载体的构建。结果表明:克隆到长252bp的完整编码基因序列,编码84个氨基酸残基;经测序与同源比对鉴定该基因与已报道基因相似性高度一致;以pCAMBIA3301为基础构建了植物表达载体pCAMBIA3301PI并转化入农杆菌EHA105中。

In order to study the safety of insecticidal protein,woth commercially available Jiyu 47 seeds as mate rials, total RNA was extracted, reverse transcription was carried out with the RNA as template, and amplifica tion J iyu 47 protoase inhibitor gene hy RT PCR, the end bioinformatics analysis and construction of expression vector were carried out. The results showed that 252 bp complete sequences was cloned,it encoded 84 amino acids; sequencing and Blast with reported gene highly similar; expression vector pCAMBIA3301PI was con strueled by pCAMBIA3301 and transformed into Agrobacterium EHA105.