苦瓜McAPX2基因及其启动子的克隆与表达分析
发布时间:2018-05-26 01:56
本文选题:苦瓜 + McAPX基因 ; 参考:《热带作物学报》2016年12期
【摘要】:基于苦瓜叶片均一化文库克隆得到McAPX2基因的cDNA全长序列(Gen Bank登录号:KJ722767.1),采用染色体步移法获得该基因的启动子序列,实时荧光定量PCR技术检测该基因在不同器官中的表达量及低温胁迫对McAPX2表达量的影响。结果表明:McAPX2基因全长1 086 bp,开放阅读框747 bp,编码249个氨基酸;McAPX2属于过氧化物酶家族ClassⅠ的成员,其脂肪族氨基酸指数为83.90,是一个亲水性蛋白,可推测定位于细胞质中;与甜瓜(NP_001284378.1)、黄瓜(ACO90193.1)、南瓜(AHF27424.1)、苦瓜(AGJ72851.1)同源性较高,分别为95%,94%,94%和93%;q RT-PCR结果显示,在苦瓜的根、茎、叶、雌花和幼果等组织器官中McAPX2的表达量存在显著差异,其中根的表达量最大,而在叶片中的表达量最低;在低温胁迫下,随着胁迫时间的延长,McAPX2表达量逐渐上调,处理12 h时达到最大,随后下降,说明该基因的表达与苦瓜耐低温胁迫相关。分离得到McAPX2基因上游调控序列1 267 bp,其启动子含有与GA、ABA、CTK、乙烯等激素和病原菌等生物胁迫以及热激、干旱、低温、光等非生物胁迫的相关元件。
[Abstract]:Based on the homogenization library of Momordica charantia L., the full-length cDNA sequence of McAPX2 gene was cloned, and the promoter sequence of the gene was obtained by chromosome step method, the accession number of GenGen Bank was: KJ722767.1. The expression of the gene in different organs and the effect of low temperature stress on the expression of McAPX2 were detected by real-time fluorescence quantitative PCR. The results showed that 1 086 BP of 1: McAPX2 gene and 747 BP of open reading frame, McAPX2 encoding 249 amino acids belong to the peroxidase family Class 鈪,
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