柠条锦鸡儿CkLEA4-1基因的克隆及表达分析

发布时间：2018-01-28 02:53

本文关键词： 柠条锦鸡儿 LEA 基因克隆表达分析　出处：《华北农学报》2017年02期 　论文类型：期刊论文

【摘要】：为研究胚胎发育晚期蛋白(LEA)在柠条中的生物学功能,从已构建的柠条锦鸡儿干旱胁迫抑制削减杂交文库中筛选到一条LEA蛋白编码基因,并采用RT-PCR法进行克隆。所得LEA基因的开放阅读框(ORF)长1 119 bp,编码373个氨基酸的蛋白质,命名为CkLEA4-1。结合序列比对与系统进化分析结果,推断CkLEA4-1属于5族LEA蛋白。利用实时荧光定量PCR技术对CkLEA4-1在干旱、高盐等逆境胁迫条件下的表达情况进行初步研究,结果表明,CkLEA4-1受到不同程度的诱导,推测该基因可能与柠条锦鸡儿响应逆境胁迫有关。研究还成功构建了CkLEA4-1的过表达载体p Can G-CkLEA4-1,得到转基因纯合体株系,为进一步研究柠条锦鸡儿CkLEA4-1基因的功能提供了材料。
[Abstract]:In order to study the biological function of Lea (late embryonic development protein) in Caragana korshinskii, a LEA protein encoding gene was screened from the constructed cDNA library of Caragana korshinskii (Caragana korshinskii). The open reading frame of LEA gene was 1 119 BP long and encoded 373 amino acids. It was named CkLEA4-1. combined with sequence alignment and phylogenetic analysis. It was inferred that CkLEA4-1 belonged to five families of LEA protein. The expression of CkLEA4-1 under stress conditions such as drought and high salt was studied by real-time fluorescence quantitative PCR. The results showed that CkLEA4-1 was induced to different degrees. It was suggested that the gene might be related to the response of Caragana korshinskii to stress stress. The overexpression vector p Can G-CkLEA4-1 of CkLEA4-1 was constructed successfully. Transgenic homozygous lines were obtained, which provided materials for further study on the function of Caragana korshinskii CkLEA4-1 gene.
【作者单位】：内蒙古农业大学生命科学学院;
【基金】：国家自然科学基金项目(31560199;31360169) 内蒙古自治区科技创新团队(201503004) 内蒙古自治区草原英才团队
【分类号】：S793.3
【正文快照】： 在植物整个生命周期中,干旱、盐碱和冻害成为限制植物生长发育的重要逆境因子,在长期的进化过程中植物为抵御这些环境胁迫,形成了特定、复杂的调控机制[1-2]。其中胚胎发育晚期蛋白(Late embryogenesis abundant,LEA)就是植物在遭遇逆境时诱导合成的一系列功能蛋白,具有保护植

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