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本研究采用RT-PCR技术克隆大豆KTi和SBA基因的核心保守序列,序列分析表明,KTi基因片段为324bp,SBA基因片段为515bp;利用RNAi技术,结合种子特异性启动子,构建同时具有KTi基因和SBA基因反向重复结构的双价RNAi种子特异性表达载体pCAMBIA3301-KTi-SBA(pKTi-SBA)。通过酶切检测及测序,证明双价RNAi种子特异性表达载体构建成功。本研究为通过RNAi技术同时降低大豆种子中胰蛋白酶抑制剂和凝集素含量,改良大豆品质奠定基础。
In this study, the core conserved sequences of KTi and SBA genes were cloned by RT-PCR. Sequence analysis showed that the KTi gene fragment was 324bp and the SBA gene fragment was 515bp. Using RNAi technology combined with seed-specific promoter, And double-stranded RNAi seed-specific expression vector pCAMBIA3301-KTi-SBA (pKTi-SBA) with inverted repeat structure of SBA gene. Through digestion detection and sequencing, the bivalent RNAi seed-specific expression vector was successfully constructed. This study laid the foundation for the improvement of soybean quality by reducing the content of trypsin inhibitor and lectin in soybean seed by RNAi technology.