黄瓜CsWRKY23基因的生物信息学及表达分析

doi:10.16035/j.issn.1001-7283.2017.05.007

摘要/Abstract

摘要：

WRKY转录因子是植物中广泛存在的一种转录因子,参与植物的生物胁迫与非生物胁迫。采用RT-PCR克隆黄瓜CsWRKY23基因cDNA序列,对序列进行结构域、基因结构分析,构建系统发育树,通过实时荧光定量PCR（qRT-PCR）分析该基因在黄瓜根、叶组织中的表达。结果表明：CsWRKY23的cDNA开放读码框全长1 431bp,含有5个外显子,4个内含子;编码476个氨基酸,含有两个WRKY结构域,编码蛋白的分子量为52.2kDa、等电点为6.43;系统发育分析表明CsWRKY23与拟南芥AtWRKY33同源。qRT-PCR结果表明该基因响应低温胁迫,并在根、叶组织中的表达均上调。本研究为进一步鉴定该基因在非生物胁迫的功能奠定了基础。

关键词: 黄瓜, WRKY, 生物信息学, 荧光定量PCR

Abstract:

WRKY transcription factor is a great gene family that exists in higher plants and widely involved in plant biotic stress and abiotic stress. The cDNA of CsWRKY23 was cloned by RT-PCR, and sequence motif, gene structure and phylogenetic tree were done. The gene expression of CsWRKY23 were detected by qRT-PCR. The results showed that the length of CsWRKY23 open reading frame was 1 431bp, containing 5 exons and 4 introns; encoding 476 amino acids, and there were 2 WRKY motifs, protein molecular weight was 52.2kDa and its iso-electric point was 6.43; phylogenetic analysis showed CsWRKY23 and AtWRKY33 were homologous. Furthermore, qRT-PCR results showed that CsWRKY23 respond to low temperature and up-regulated in leaf and root. In addition, this study provided useful information for further studying this gene and its function under the abiotic stress.

Key words: Cucumber, WRKY, Bioinformatics, Quantitive real time PCR

张颖,刘朋宇,白雪,杨阳,李玥莹. 黄瓜CsWRKY23基因的生物信息学及表达分析[J]. 作物杂志, 2017 (5): 38–42

Ying Zhang,Pengyu Liu,Xue Bai,Yang Yang,Yueying Li. Expression and Bioinformatics Analysis of CsWRKY23 Gene in Cucumber[J]. Crops, 2017(5): 38–42

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序号Code	元件Element	位置Location	序列Sequence	功能Function
1	WRKY71OS	47	TGAC	WRKY转录因子的结合位点
2	TATABOXOSPAL	74	TATTTAA	转录起始位点上游通用转录因子结合的主要的启动原件
3	WRKY71OS	98	TGAC	WRKY转录因子的结合位点
4	CAATBOX1	166	CAAT	增强结合蛋白因子
5	CBFHV	225	RYCGAC	响应低温的CBF转录因子,AP2结构域的结合位点
6	MYBCORE	530	CNGTTR	MYB转录因子结合位点
7	WRKY71OS	584	TGAC	WRKY转录因子的结合位点
8	WRKY71OS	1386	TGAC	WRKY转录因子的结合位点
9	ABREATCONSENSUS	1668	YACGTGGC	ABA响应过程中的顺势作用原件
10	WRKY71OS	1721	TGAC	WRKY转录因子的结合位点
11	WRKY71OS	1749	TGAC	WRKY转录因子的结合位点
12	WRKY71OS	1977	TGAC	WRKY转录因子的结合位点

基因Gene	登录号Accession number	引物序列Primer sequence	用途Utility
CsWRKY23	GU984022	F:ATGAGTAACATAAACCAAAC R:CTATGATAGAAATGAGTTG	cDNA克隆全长引物
CsWRKY23	GU984022	F:CACTACCTACGAAGGCAAAC R:TGGCTGGTGACACTGGAT	qRT-PCR引物
CsActin	AAZ74666	F:TCCACGAGACTACCTACAACTC R:CGGAATGGTGAAGGCTGGAT	qRT-PCR引物