RinPKS1基因过表达载体的构建及遗传转化树莓

摘要

RinPKS 1 encodes a bifunctional enzyme having both CHS activity and BAS activity that can catalyze the synthesis of naringin and benzalacetone.BAS is also the key enzyme involved in the biosynthesis of raspberry ketone.[Objective]To verify whether RinPKS 1 play a role in the synthesis of raspberry ketone.[Methods] In this study,we generated PCA-RinPKS 1 constructed by ligation RinPKS 1 with pCambia 1304 and then transformed into the raspberry by using Agrobacterium tumefaciens.The expression level of RinPKS 1 was detected using q-PCR.[Results] The results showed that the expression levels of RinPKS 1 in root,stem and leaf of the PCA-RinPKS 1 transformed raspberry are 15.8 folds,3.17 folds and 4.39 folds,respectively higher than that of the negative control.[Conclusion]RinPKS 1 has overexpressed in transcriptional level in raspberry.%RinPKS 1基因编码的蛋白是具有催化合成柚皮素查尔酮和苯亚甲基丙酮的能力双功能酶,即具有CHS和BAS活性.BAS又是树莓酮生物合成途径中的关键酶.[目的]为了验证RinPKS 1在树莓酮生物合成途径中是否发挥作用.[方法]将RinPKS 1基因与pCambia 1304连接,构建成植物过表达载体PCA-RinPKS1,通过根癌农杆菌介导,将RinPKS 1遗传转化入树莓中,以提高树莓中树莓酮的含量.利用q-PCR分别检测转PCA-RinPKS1和阴性对照(空载体)的树莓材料中RinPKS 1基因的表达量.[结果]转RinPKS 1基因树莓材料中RinPKS 1的表达量比转阴性对照(空载体)有很大提高,分别是根高出15.8倍;茎高出3.17倍;叶高出4.39倍.[结论]RinPKS 1在树莓中得到转录水平过表达.