火龙果己糖转运蛋白基因克隆及生物信息学分析

    Cloning and Bioinformatics Analysis of A Hexose Transporter Gene in Pitaya

    • 摘要:
      目的 己糖转运蛋白(HexoseTransporter,HXT)是一种单糖转运蛋白,能够跨膜转运甘露糖、果糖和葡萄糖等己糖类物质,参与花粉发育和植株生长,进而决定果实的内在品质。分析火龙果HXT基因生物信息学及表达特性,可为调节火龙果果实品质奠定基础。
      方法 通过基因克隆得到1个火龙果己糖转运蛋白基因,应用NCBI、BioEdit和DNAMAN等软件或数据库对其进行生物信息学分析,并利用HLPC技术分析成熟期果实中的可溶性糖含量,采用荧光定量PCR等方法分析不同株系火龙果及果实中不同部位的HTX基因的表达特性。
      结果 通过PCR扩增从火龙果中克隆得到1个HXT基因,命名为HuHXT。该基因序列全长1 230 bp,编码410个氨基酸。蛋白不稳定系数35.59,有32个Ser、15个Thr和15个Tyr。火龙果中可溶性糖组分主要为蔗糖、葡萄糖和果糖,以葡萄糖和果糖为主。HuHXT在97号株系中表达量最高,其次为湛红2号(ZH)和128号株系,15号和16号株系表达量较少;HuHXT的表达量在同一果实不同部位差别不显著,在靠近果皮部位表达量较高。
      结论 HuHXT为Sugar-tr基因家族成员,该蛋白属于稳定蛋白,疏水性较强,是一种膜蛋白,二级结构以α- 螺旋和无规则卷曲为主。HuHXT表达量与果实中葡萄糖含量呈负相关关系,即HuHXT表达量越高、葡萄糖含量越少。

       

      Abstract:
      Objective  Hexose transporter (HXT) is a kind of monosaccharide transporter, which can transport mannose, fructose, glucose and other hexose substances across the membrane, and participate in the development of pollen and the growth of the whole plant, and then determine the internal quality of fruit. Bioinformatics and expression characteristics of HXT gene in pitaya were analyzed in order to provide a theoretical basis for regulating the fruit quality of pitaya.
      Method  A hexose transporter gene was obtained by gene cloning. Bioinformatics analysis was conducted by using NCBI, BioEdit, DNAMAN and other software or databases. The soluble sugar contents were determined by HPLC at fruit maturity stage. The relative expression characteristics of HTX gene in different lines of pitaya and different parts of afruit were analyzed by fluorescence quantitative PCR.
      Result  A HXT gene named HuHXTwas cloned from pitaya fruit by PCR amplification. The total length of the gene was 1 230 bp, encoding 410 amino acids. The protein instability coefficient was 35.59, with 32 Ser, 15 Thr and 15 Tyr. There were sucrose, glucose and fructose in pitaya, and glucose and fructose were the main components. The expression of HuHXT was the highest in No. 97 line, followed by Zhanhong No. 2 and No. 128, while No. 15 and No. 16 had the lowest expression level. The expression level of HuHXT was not significantly different in different parts of a fruit. The expression in parts near the pericarp was higher other parts of a fruit.
      Conclusion  HuHXT is a member of the Sugar-tr gene family. HuHXT protein is a stable protein with strong hydrophobicity. As a membrane protein, its secondary structure is mainly α-helix and random curl. There is negative correlation between the relative expression of HuHXT and the content of glucose, that is, the higher the HuHXT expression level, the lower the glucose content.

       

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