玉米<i>ZmTOC1a、ZmTOC1b</i>基因的克隆、表达及亚细胞定位分析

doi:10.7668/hbnxb.201751601

摘要/Abstract

摘要： 拟南芥TOC1基因在拟南芥中与2个MYB类蛋白基因LHY（Late elongated hypocotyl）、CCA1（Circadian clock associated1）组成中央振荡器，通过光周期调节途径调控拟南芥对光照的响应。为了揭示玉米中央振荡器中重要基因ZmTOC1a与ZmTOC1b的生物学功能，通过同源搜索找到玉米中的2个同源基因ZmTOC1a与ZmTOC1b，并通过同源克隆得到了这2个基因的序列，进而对这2个基因进行组织表达分析和编码蛋白的亚细胞定位。结果表明，通过同源克隆得到ZmTOC1a的开放阅读框的总长度为1 236 bp，共编码411个氨基酸；ZmTOC1b的开放阅读框的全长为1 554 bp，共编码517个氨基酸。通过ProtPARAM进行基因编码蛋白质的理化性质分析，ZmTOC1a与ZmTOC1b均为酸性蛋白。NetPhos 3.1 Server预测结果显示，ZmTOC1a存在46个潜在磷酸化位点，其中丝氨酸36个，苏氨酸8个，酪氨酸2个；ZmTOC1b共存在53个潜在磷酸化位点，丝氨酸38个，苏氨酸12个，酪氨酸3个。在玉米中选取11个不同的组织进行qRT-PCR分析，结果表明，ZmTOC1a及ZmTOC1b分别在胚根以及胚芽鞘中高表达。通过构建目的蛋白与GFP融合的表达载体并注射烟草，发现ZmTOC1a及ZmTOC1b表达蛋白主要集中在细胞核中。综上所述，玉米TOC1基因可能与拟南芥TOC1基因作用一致，在玉米的生物钟调节中起到了重要的作用。

关键词: 玉米, ZmTOC1a, ZmTOC1b, 克隆, 组织表达分析, 亚细胞定位

Abstract: The Arabidopsis TOC1 gene, together with two MYB protein gene LHY (Late elongated hypocotyl)and CCA1 (Circadian clock associated1), constitutes the central oscillator, which regulates the growth of Arabidopsis through photoperiod pathway. In order to explore the biological functions of ZmTOC1a and ZmTOC1b in central oscillator of maize, by means of homology search, two homologous genes of ZmTOC1a and ZmTOC1b from maize were detected, and then were cloned. The expressions patterns and subcellular localization of ZmTOC1a and ZmTOC1b were conducted. The results showed that the full length of ZmTOC1a was 1 236 bp, encoding 411 amino acids. The ORF length of ZmTOC1b was 1 554 bp, encoding 517 amino acids. ProPARAM analysis showed that the physical and chemical properties of ZmTOC1a and ZmTOC1b proteins were acidic. The predicted results of NetPhos 3.1 Server indicated that ZmTOC1a had 46 Phosphorylation sites, including 36 serine, 8 threonine and 2 tyrosine sites. ZmTOC1b had 53 phosphorylation site, including 38 serine, 12 threonine and 3 tyrosine. According to the results of qRT-PCR in different organs, the expression levels of ZmTOC1a and ZmTOC1b were the highest in radicle and coleoptile, respectively. Furthermore, the subcellular localization showed that the expression of ZmTOC1a and ZmTOC1b was observed mainly in nucleus. Above all, the TOC1 genes might play an important role in photoperiod pathway of maize.

Key words: Maize, ZmTOC1a, ZmTOC1b, Clone, Tissue expression analysis, Subcellular localization

中图分类号:

Q78
S513.03

蔡云婷, 贾力, 拓昊苑. 玉米ZmTOC1a、ZmTOC1b基因的克隆、表达及亚细胞定位分析[J]. 华北农学报, 2019, 34(4): 24-31. doi: 10.7668/hbnxb.201751601.

CAI Yunting, JIA Li, TUO Haoyuan. Cloning, Expression and Subcellular Localization of ZmTOC1a and ZmTOC1b Genes in Maize[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(4): 24-31. doi: 10.7668/hbnxb.201751601.

http://www.hbnxb.net/CN/Y2019/V34/I4/24

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