紫花苜蓿MsSNARE基因的克隆及表达分析
发布时间：2018-07-13 17:32
【摘要】：紫花苜蓿(Medicago sativa L.),是一种栽培范围广、适应性强、品质好、产量高的豆科牧草。虽然自身具有很强的非生物胁迫抗性,但全球气候变化及极端天气的发生,依然会给苜蓿生产造成严重损失。因此,培育具有优良抗性的紫花苜蓿品种十分重要。近年来,关于紫花苜蓿相关基因的克隆、功能验证及遗传转化的研究进展较快。植物中的SNARE蛋白可以提高植物的抗病性,通过与其他通道蛋白的互作影响植物的生长发育及对环境刺激的应答。本研究利用RACE法克隆了紫花苜蓿MsSNARE基因,并利用生物信息学软件对其序列进行分析;构建了瞬时表达载体PBI-MsSNARE;通过qRT-PCR技术对该基因的表达模式进行分析。主要研究结果如下:1.利用RACE法,从紫花苜蓿中成功克隆出MsSNARE基因cDNA的全长,经序列分析并发现MsSNARE cDNA全长1396 bp,开放阅读框长(ORF)1257 bp,共编码418个氨基酸残基,其编码的蛋白属于SNARE家族。GeneBank登录号为:JX467688。2.qRT-PCR分析发现MsSNARE基因在紫花苜蓿的根、茎、叶中都有表达,在根、茎和叶中的表达量差异性不显著。3.qRT-PCR分析了MsSNARE基因在受到干旱、盐胁迫、ABA诱导时的转录水平变化,结果发现在受到外界胁迫时MsSNARE表达水平均上调。4.构建了MsSNARE基因瞬时表达载体PBI-MsSNARE,洋葱表皮细胞亚细胞定位研究表明该基因的产物定位于细胞膜上。
[Abstract]:Alfalfa (Medicago sativa L.) is a leguminous forage with wide cultivation range, strong adaptability, good quality and high yield. Although it has strong abiotic stress resistance, the global climate change and the occurrence of extreme weather will still cause serious losses to alfalfa production. Therefore, it is very important to cultivate alfalfa varieties with good resistance. In recent years, studies on cloning, functional verification and genetic transformation of alfalfa related genes have developed rapidly. The SNARE protein in plants can improve plant disease resistance and affect the growth and development of plants and their responses to environmental stimuli through interaction with other channel proteins. In this study, the MsSNARE gene of alfalfa was cloned by race method, and its sequence was analyzed by bioinformatics software. The transient expression vector PBI-MsSNAREwas constructed, and the expression pattern of the gene was analyzed by qRT-PCR. The main results are as follows: 1. The full length of MsSNARE gene cDNA was cloned from alfalfa by race method. By sequence analysis, the full length of MsSNARE cDNA was found to be 1396 BP, and the open reading frame length (ORF) was 1257 BP, encoding 418 amino acid residues. The encoding protein belongs to the Snare family. The accession number of GeneBank is: JX467688.2.qRT-PCR analysis shows that the MsSNARE gene is expressed in the roots, stems and leaves of alfalfa, but there is no significant difference in the expression levels in the roots, stems and leaves. 3.qRT-PCR analysis showed that the MsSNARE gene was subjected to drought. The results showed that the expression level of MsSNARE was up-regulated under the stress of salt stress, and the results showed that the expression level of MsSNARE was up-regulated by .4. The transient expression vector PBI-MsSNAREof MsSNARE gene was constructed. The subcellular localization of MsSNARE gene in epidermal cells of onion showed that the product of MsSNARE gene was located on the cell membrane.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S541.9
本文编号：2120186
[Abstract]:Alfalfa (Medicago sativa L.) is a leguminous forage with wide cultivation range, strong adaptability, good quality and high yield. Although it has strong abiotic stress resistance, the global climate change and the occurrence of extreme weather will still cause serious losses to alfalfa production. Therefore, it is very important to cultivate alfalfa varieties with good resistance. In recent years, studies on cloning, functional verification and genetic transformation of alfalfa related genes have developed rapidly. The SNARE protein in plants can improve plant disease resistance and affect the growth and development of plants and their responses to environmental stimuli through interaction with other channel proteins. In this study, the MsSNARE gene of alfalfa was cloned by race method, and its sequence was analyzed by bioinformatics software. The transient expression vector PBI-MsSNAREwas constructed, and the expression pattern of the gene was analyzed by qRT-PCR. The main results are as follows: 1. The full length of MsSNARE gene cDNA was cloned from alfalfa by race method. By sequence analysis, the full length of MsSNARE cDNA was found to be 1396 BP, and the open reading frame length (ORF) was 1257 BP, encoding 418 amino acid residues. The encoding protein belongs to the Snare family. The accession number of GeneBank is: JX467688.2.qRT-PCR analysis shows that the MsSNARE gene is expressed in the roots, stems and leaves of alfalfa, but there is no significant difference in the expression levels in the roots, stems and leaves. 3.qRT-PCR analysis showed that the MsSNARE gene was subjected to drought. The results showed that the expression level of MsSNARE was up-regulated under the stress of salt stress, and the results showed that the expression level of MsSNARE was up-regulated by .4. The transient expression vector PBI-MsSNAREof MsSNARE gene was constructed. The subcellular localization of MsSNARE gene in epidermal cells of onion showed that the product of MsSNARE gene was located on the cell membrane.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S541.9
【相似文献】
相关硕士学位论文 前1条
1 张通;紫花苜蓿MsSNARE基因的克隆及表达分析[D];西北农林科技大学;2016年
，本文编号：2120186
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2120186.html
最近更新
教材专著
