盐生植物四翅滨藜Actin基因片段的克隆及表达分析

发布时间：2018-06-01 12:28

本文选题：四翅滨藜 + 肌动蛋白　；参考：《草业科学》2017年03期

【摘要】：在研究植物功能基因表达模式中肌动蛋白(Actin)编码基因是最常用的内参基因之一。本研究根据藜科植物Actin基因编码区保守序列设计一对简并引物,以盐生植物四翅滨藜(Atriplex canescens)4周龄幼苗叶片的总RNA为模板,采用逆转录PCR(reverse transcription PCR,RT-PCR)方法扩增出Actin基因片段并连接到pMD19-T载体上,所得阳性克隆经PCR检测后进行测序。序列分析结果表明,该基因片段长度为598bp,编码198个氨基酸;该序列与Genbank中登录的高等植物Actin基因核苷酸序列同源性在84%以上,氨基酸序列同源性在94%以上。说明其为Actin基因片段,具有高度保守性,命名为AcACT。序列进化分析显示,AcACT与甜菜(Beta vulgaris)和碱蓬(Suaeda glauca)肌动蛋白编码基因的亲缘关系最近。荧光定量PCR分析结果表明,AcACT在盐处理下各组织中的表达量恒定。表明AcACT可作为内参基因来研究四翅滨藜耐盐相关基因的表达模式。
[Abstract]:Actin (actin) coding gene is one of the most common internal reference genes in studying the expression pattern of plant functional genes. In this study, a pair of degenerate primers were designed according to the conserved sequence of Actin gene coding region in Chenopodiaceae. The total RNA of Atriplex canescens)4 week-old seedling leaves of halophytes was used as template. The Actin gene fragment was amplified by reverse transcription PCR(reverse transcription PCR RT PCR and ligated to the pMD19-T vector. The positive clones were sequenced after PCR detection. The sequence analysis showed that the length of the gene was 598 BP, encoding 198 amino acids, and the nucleotide sequence homology was over 84% and the amino acid sequence homology was over 94% with the Actin gene of higher plants registered in Genbank. It is a highly conserved Actin gene fragment named Acac. Sequence analysis showed that AcACT was closely related to the actin coding genes of Beta vulgaris and Suaeda glauca. Fluorescence quantitative PCR analysis showed that the expression of AcACT in salt treated tissues was constant. The results indicated that AcACT could be used as an internal reference gene to study the expression pattern of salt-tolerant genes in Chenopodium angustifolia.
【作者单位】：兰州大学草地农业科技学院草地农业生态系国家重点实验室;
【基金】：国家自然科学基金(31372360) “本科教学工程”国家级大学生创新创业训练计划项目(201610730105) “中央高校基本科研业务费专项资金”自由探索项目(lzujbky-2015-250)
【分类号】：Q943.2

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