| 李甜子，郑雪文，曹新玥，陈嘉华，杨转英.火龙果己糖转运蛋白基因克隆及生物信息学分析[J].广东农业科学,2022,49(3):31-39 |
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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 |
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| DOI：10.16768/j.issn.1004-874X.2022.03.004 |
| 中文关键词: 火龙果 己糖转运蛋白 生物信息学 葡萄糖 基因表达 |
| 英文关键词: pitaya hexose transporter bioinformatics glucose gene expression |
| 基金项目:广东海洋大学大学生创新创业项目（230429299）；广东省农村科技特派员驻镇帮镇扶村项目（A21411）；国家重点研发计划项目（2019YFD1002205） |
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| 中文摘要: |
| 【目的】己糖转运蛋白（Hexose Transporter，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。 |
| 英文摘要: |
| 【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 a fruit were analyzed by fluorescence quantitative PCR. 【Result】A HXT gene named HuHXT was 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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