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以油桐转录组数据中FATB1基因的cDNA片段为模板,通过RACE技术克隆获得该基因的CDS,命名为Vf FATB1,并进行了生物信息学分析,同时通过Real-time PCR和亚细胞定位对该基因的表达模式和表达部位进行了研究分析。结果表明Vf FATB1是一个由8个α-螺旋和10个β-折叠组成“hot-dog”折叠结构的二聚体,与JcFATB1相似度高达92%;该基因有FAT重要催化作用的半胱氨酸活性位点Cys320和3个保守的组氨酸位点His140/285/345;该基因在油脂合成后期种仁中的表达量最高,是幼根的30倍左右,且在油脂合成过程中一直是上调状态;该基因定位于叶绿体中。综上所述,Vf FATB1是一个位于叶绿体中且含有多个催化功能结构的蛋白,表达模式与油脂积累相同,因此它可能与油脂合成有着密切的关系,这为油桐的遗传改良提供了潜在的基因资源。
The cDNA fragment of FATB1 gene in Tung oilseed transcriptome data was used as a template and the gene CDS was cloned by RACE technology and named as Vf FATB1. Bioinformatic analysis was also carried out. The real-time PCR and subcellular localization Gene expression patterns and expression sites were studied. The results showed that Vf FATB1 is a dimer composed of “hot-dog” folding structure consisting of 8 α-helices and 10 β-sheets and has a similarity of 92% with JcFATB1. This gene has half of the important catalytic effects of FAT Cystine active site Cys320 and three conserved histidine His140 / 285/345 sites; the gene expression in the kernel during late oil synthesis is the highest, about 30 times the young root, and in the oil synthesis process Has been up-regulated; the gene is localized in the chloroplast. Taken together, Vf FATB1, a chloroplast-containing protein with multiple catalytic structures, expresses the same pattern of lipid accumulation. Therefore, it may be closely related to lipid synthesis, which provides potential for genetic improvement of tung tree The genetic resources.