越橘谷胱甘肽巯基转移酶基因的克隆及表达

发布时间：2018-02-09 20:35

本文关键词： 越橘花青素谷胱甘肽巯基转移酶基因克隆基因表达　出处：《吉林农业大学学报》2017年04期 　论文类型：期刊论文

【摘要】：以越橘(Vaccinium L.)品种"北陆"为试材,采用反转录PCR克隆技术成功克隆越橘谷胱甘肽巯基转移酶基因c DNA全长序列,获得一个完整的编码区c DNA序列,长为690 bp,编码229个氨基酸,将其命名为VcGSTU1(Gen Bank登录号KT601064)。经与谷胱甘肽巯基转移酶家族其他成员的氨基酸序列比对和系统进化分析,初步确定VcGSTU1属于谷胱甘肽巯基转移酶Tau家族。生物信息学分析表明,该蛋白不稳定指数为28.80,属于稳定蛋白。预测该蛋白分子量为25.499 ku,等电位点为5.68。蛋白质序列同源比对发现其含有完整的GST-N-Tau和GST-C-Tau结构域。利用反转录PCR和实时荧光定量PCR(qRT-PCR)2种方法分析谷胱甘肽巯基转移酶的表达模式,该基因在越橘的根、茎、叶芽、叶、花芽、绿果、粉果、蓝果、蓝果果皮、蓝果种子中均有表达,且在蓝果种子中相对表达量最高。
[Abstract]:The full-length c DNA sequence of glutathione mercaptotransferase gene (c DNA) was cloned successfully by reverse transcription PCR cloning from "Beilu", a variety of blueberry Vaccinium L., and a complete coding region c DNA was obtained, which was 690 BP long and encoded 229 amino acids. It was named VcGSTU1(Gen Bank accession number KT601064. After amino acid sequence alignment and phylogenetic analysis with other members of glutathione mercaptotransferase family, it was preliminarily confirmed that VcGSTU1 belongs to the glutathione mercaptotransferase Tau family. Bioinformatics analysis showed that VcGSTU1 belonged to glutathione mercaptotransferase Tau family. The instability index of the protein is 28.80, which belongs to the stable protein. The predicted molecular weight of the protein is 25.499ku. the equipotential point is 5.68.The homologous alignment of the protein sequence shows that the protein contains complete GST-N-Tau and GST-C-Tau domains. Reverse transcription PCR and real-time fluorescence are used to determine the protein. The expression pattern of glutathione mercaptotransferase was analyzed by quantitative PCR(qRT-PCR)2 method. The gene was expressed in root, stem, leaf bud, leaf, flower bud, green fruit, powdery fruit, blue fruit, blue fruit peel and blue fruit seed of blueberry, and the highest relative expression was found in blueberry seed.
【作者单位】：吉林农业大学园艺学院;
【基金】：吉林省科技发展计划项目(20150414047GH) 吉林省省级经济结构战略调整引导资金专项(2015Y060) 长春市农业先进实用技术示范推广计划项目(14NK010)
【分类号】：S663.9

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