海岛棉GbTCP14基因的克隆与表达分析
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国家自然科学基金(31560405);


Cloning and Expression Analysis of GbTCP14 Gene in Gossypium barbadense
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    摘要:

    以海岛棉‘新海21号’为试验材料,根据陆地棉GhTCP14基因序列,设计1对引物,通过RTPCR技术获得海岛棉GbTCP14核苷酸序列,开放阅读框长1 221 bp,编码406个氨基酸,分子式C1892H2950N572O620S16,预测分子量为44.134 6 kD,等电点为6.88,氨基酸序列包含1个高度保守的TCP结构域及4个低丰度复杂区。GbTCP14蛋白氨基酸序列与其他物种TCP14氨基酸序列比对表明,海岛棉GbTCP14蛋白与其他植物中的TCP14蛋白共同具有一个高度保守的TCP结构域,序列之间的一致性较高。进化树分析表明,海岛棉GbTCP14基因与木本棉GaTCP14基因分布在同一分支上。实时荧光定量PCR表明,海岛棉GbTCP14基因在开花后第15天时表达量最高,在第5~20天时相对表达量比其他时期较高,第25天时相对表达量最低。在不同组织中花瓣和花萼中表达量较高,根和茎中表达量一般,叶组织中表达量最低。通过酵母系统转录激活分析显示,GbTCP14蛋白不具有转录活性。

    Abstract:

    The ‘New Sea 21’ of Gossypium barbadense was used as the experimental material. A pair of primers were designed according to the GhTCP14 gene sequence of upland cotton. A nucleotide sequence was obtained by RTPCR. The open reading frame was 1 221 bp, encoding 406 amino acids, molecular formula C1892H2950N572O620S16, the predicted molecular weight was 44.134 6 kD, the isoelectric point was 6.88, and the amino acid sequence contained one highly conserved TCP domain and four low abundance complex regions. The comparison of amino acid sequence of GbTCP14 protein with other species TCP14 showed that the GbTCP14 protein of G. barbadense had a highly conserved TCP domain with the TCP14 protein in other plants, and the consistency between the sequences is high. The results of phylogenetic tree showed that the GbTCP14 gene of G. barbadense was distributed on the same branch as Grafted cotton GaTCP14 gene. Realtime quantitative PCR showed that the expression level of GbTCP14 gene was the highest at the 15th day after flowering, and the relative expression level was higher on the 5th day to the 20th day than that in other days. The expression level in petals and calyx was higher among different tissues, and the expression level in roots and stems was the lowest. GbTCP14 protein did not have transcriptional activity by yeast system transcriptional activation assays. The results showed that this study successfully cloned a TCP transcription factor from the island of cotton. The preliminary analysis of the gene has a good potential capacity, the specific function to be further studied.

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蔡永生,郑 凯,王 怡,等.海岛棉GbTCP14基因的克隆与表达分析[J].西北植物学报,2018,38(3):401-408

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  • 在线发布日期: 2018-04-23
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