菊花叶绿素a/b结合蛋白基因CmLhcb1及其启动子的克隆和表达分析

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以切花菊品种‘公子’cDNA为模板克隆出叶绿素a/b结合蛋白同源基因cab,其开放阅读框为798bp,编码266个氨基酸。经多物种间比对分析,确认其属于cab基因家族的Lhcb1类,命名为CmLhcb1。同源克隆菊花品种‘清露’Lhcb1基因,氨基酸序列与‘公子’完全相同。CmLhcb1在叶片中的表达量比在茎、花和根中高,弱光和GA3处理使CmLhcb1表达上调,多效唑处理后CmLhcb1表达量受到抑制。CmLhcb1的表达受昼夜节律调节,白天表达量显著高于夜间。通过high-efficiencyTAIL-PCR(hiTAIL-PCR)方法克隆到‘公子’切花菊CmLhcb1起始密码子上游序列715bp和‘清露’起始密码子上游序列716bp,序列经PLACE数据库的比对分析,发现有很多与非生物和生物胁迫相关的元件,主要与光照、GA、ABA、水分、水杨酸和病毒相关,CmLhcb1启动子是光诱导型启动子,具有GT1-box和Z-box。 The chlorophyll a / b binding protein homologous gene cab was cloned using the cut chrysanthemum cDNA of ’Son’ as its template. Its open reading frame is 798 bp, encoding 266 amino acids. According to the cross-species alignment analysis, it was confirmed that it belonged to the Lhcb1 class of cab gene family and named CmLhcb1. The homologous cloned chrysanthemum variety ’Qinglu’ Lhcb1 gene, the amino acid sequence and ’son’ exactly the same. The expression level of CmLhcb1 in leaves was higher than that in stems, flowers and roots, and the expression of CmLhcb1 was up-regulated by low light and GA3 treatment. The expression of CmLhcb1 was inhibited after paclobutrazol treatment. The expression of CmLhcb1 was regulated by the circadian rhythm and the expression level during daytime was significantly higher than that at night. The sequence of 715bp upstream of the start codon of CmLhcb1 and 716bp of the start codon of ’Qinglu’ was cloned by high-efficiency TAIL-PCR (hiTAIL-PCR) method and the sequence was analyzed by PLACE database. There are many elements involved in abiotic and biotic stress, mainly related to light, GA, ABA, moisture, salicylic acid and viruses. The CmLhcb1 promoter is a light-inducible promoter with GT1-box and Z-box.
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