羊草14-3-3基因的克隆及序列分析  

Isolation and Expression Analysis of 14-3-3 Gene fromLeymus chinensis

作　　者：赵英 刘晶莹[2] 杨梦丹 汪自庆[1] 麻鹏达[2] 

机构地区：[1]西北农林科技大学理学院,陕西杨凌712100 [2]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《西北农业学报》2018年第2期269-274,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31670295);中央高校基本科研业务费专项(2452017159)~~

摘　　要：通过羊草(Leymus chinensis)EST(Expressed sequence tags,EST)序列和RACE(Rapid-amplification of cDNA ends)技术,获得羊草14-3-3蛋白的全长基因为1 048bp,具有792bp开放读码框(Open reading frame,ORF)、81bp的5′非编码区(Untranslated regions,UTR)和175bp的3′UTR,编码含有263个氨基酸的蛋白质。通过序列比对,羊草14-3-3蛋白与其他物种14-3-3蛋白相似度达到80%~99%。通过RT-PCR(Reverse transcription-PCR)分析表明羊草14-3-3基因在叶片和根部均有表达,且叶片的表达量明显高于根部,而经过NaCl、Na2CO3和PEG的胁迫处理,羊草叶片14-3-3基因表达量无明显变化。本研究有助于了解14-3-3蛋白家族基因的功能,为进一步应用该基因进行植物的耐盐性遗传改良奠定基础。The 14-3-3 protein is a family of regulatory proteins encoded by agene.Although genomewide analysis of this family has been carried out in some plant species,little is known about 14-3-3 genes in Leymus chinensis.In this study,the full-length gene of Leymus chinensis14-3-3 protein was obtained by EST and RACE.The gene was 1 048 bp in length with an ORF of 792 bp,a 5′-untranlstaed region（UTR）of 81 bp and a 3′UTR of 175 bp,which encoded a predicted polypeptide of 263 amino acids.By sequence alignment,the similarity of 14-3-3 protein of Leymus chinensis and other species 14-3-3 protein was 80%-99%.The results of reverse transcription-PCR showed that Leymus chinensis 14-3-3 protein was expressed in leaves and roots,and the contents of leaves were higher than those of roots.Tracing with NaCl,Na2 CO3 and PEG 6000,the content of Leymus chinensis 14-3-3 protein did not change significantly.This study is helpful to understand the function of 14-3-3 protein family gene,which can lay the foundation for further application of this gene for salt tolerance genetic improvement inplant.

关 键 词：14-3-3蛋白 羊草 RACE RT-PCR 

分 类 号：Q785[生物学—分子生物学]