| 黄瓜酪氨酸硫化转移酶基因CsTPST克隆及其表达分析 |
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| 引用本文： | 杜亚琳.黄瓜酪氨酸硫化转移酶基因CsTPST克隆及其表达分析[J].南方农业学报,2016(9):1457-1462. |
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| 作者姓名： | 杜亚琳 |
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| 作者单位： | 农业部东北地区园艺作物生物学与种质创制重点实验室/东北农业大学 园艺学院,哈尔滨,150030 |
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| 基金项目： | 黑龙江省青年科学基金项目(QC2010089) |
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| 摘 要： | 【目的】克隆黄瓜酪氨酸硫化转移酶基因(CsTPST),并分析植物多肽在发育过程中的调控作用,为研究CsTPST的功能及多肽与乙烯在黄瓜发育过程中的相互作用打下基础。【方法】以拟南芥TPST蛋白序列信息为基础进行同源比对,经PCR扩增和生物信息学分析获得CsTPST基因序列信息、结构和亲缘关系,并利用qRT-PCR对其表达模式进行分析。【结果】CsTPST基因cDNA全长789 bp,编码262个氨基酸。进化分析结果表明,CsTPST蛋白与甜瓜的TPST蛋白亲缘关系最近,其氨基酸序列相似性为93%。qRT-PCR分析结果表明,CsTPST基因在黄瓜雌花、根和叶片中表达量较高,在雄花和茎中表达量较低。此外,CsTPST基因在乙烯合成促进剂ACC处理的黄瓜叶片中上调表达,而在乙烯合成抑制剂AVG处理下呈下调表达。CsTPST基因启动子能激活GFP基因的表达。【结论】CsTSPT基因是一个受乙烯诱导表达的基因,可供开展CsTPST基因的生物学功能及性别决定的分子机理研究参考。
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| 关 键 词： | 黄瓜 CsTPST基因 多肽 qRT-PCR 乙烯 |
Cloning and expression analysis of tyrosylprotein sulfotransferase gene CsTPST in Cucumis sativus L. |
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| Abstract: | Objective]The present study was conducted to clone tyrosylprotein Sulfotransferases gene in Cucumis sativus L.(CsTPST) and analyze regulatory role of plant peptides in development of C. sativus L., in order to lay a founda-tion for analysis of function of CsTPST and interaction between peptide and ethylene in development of C. sativus L.Method]Homology alignment was conducted based on TPST protein sequence in Arabidopsis thaliana. sequence, gene structure,phylogenetic relationship,and expression pattern of CsTPST were obtained through PCR amplification and bioin-formatics analysis, and the expression pattern was studies through qRT-PCR. Result]Gene cDNA in CsTPST was 789 bp in length,and encoded a protein with 262 amino acids. Phylogenetic analysis indicated that CsTPST exhibited the highest similarity with TPST from melon,with 93% of amino acid sequence similarity. qRT-PCR analysis showed that CsTPST gene had high expression level in female flowers,roots and leaves,while it had relatively low expression level in male flowers and stems. More interestingly,CsTSPT gene expression was up-related in cucumber leaves under ACC(an ethylene synthesis promoter) treatment,while down-regulated under AVG (an ethylene synthesis inhibitor) treatment. CsTPST gene pro-moter could activate expression of GFP gene. Conclusion]The results revealed that CsTSPT is an ethylene-induced gene, which can provide reference for research on biological function and molecular mechanism of sex determination. |
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| Keywords: | Cucumis sativus L CsTPST gene peptide qRT-PCR ethylene |
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