干旱区生态环境知识资源中心(Arid): 荒漠昆虫小胸鳖甲Ras GTP酶激活蛋白基因MpRasGAP的克隆及低温表达分析

Arid

	荒漠昆虫小胸鳖甲Ras GTP酶激活蛋白基因MpRasGAP的克隆及低温表达分析
其他题名	Cloning and expression profiling in response to low temperature of Ras GTPase-activating protein gene MpRasGAP in the desert beetle Microdera punctipennis (Coleoptera: Tenebrionidae)
	阮梦鸽; 李洁琼; 孟闪闪; 马纪
来源期刊	昆虫学报
ISSN	0454-6296
出版年	2015
卷号	58 期号:4 页码:367-374
中文摘要	[目的]丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)级联是细胞的重要信息传递系统之一,Ras GTP酶激活蛋白(Ras GTPase-activating protein,Ras GAP)基因Ras GAP和c-Jun氨基末端激酶(c-Jun Nterminal kinase,JNK)基因JNK分别是MAPK信号转导途径的上、下游基因。本研究旨在确定荒漠昆虫小胸鳖甲Microdera punctipennis Ras GAP及JNK基因对低温的响应情况。[方法]从荒漠甲虫小胸鳖甲中克隆获得Ras GAP基因的c DNA序列,利用生物信息学分析软件分析其氨基酸序列并构建进化树,利用实时荧光定量PCR检测低温胁迫条件下Ras GAP和JNK基因的表达情况。[结果]小胸鳖甲Ras GAP c DNA的开放阅读框2 523 bp,命名为MpRas GAP(Gen Bank登录号:KM677930),编码840个氨基酸,分子量96.594 k Da,编码蛋白MpRas GAP属于Ras GAP超家族。MpRas GAP与赤拟谷盗Tribolium castaneum Ras GAP的氨基酸序列一致性达89%。小胸鳖甲在4℃和-4℃低温胁迫1 h后,MpRas GAP的mRNA水平都显著高于室温对照(25℃)。小胸鳖甲在4℃处理3 h或-4℃处理1 h后,Mp JNK的mRNA水平也显著升高。[结论]本研究结果表明小胸鳖甲MpRas GAP和Mp JNK的mRNA水平受低温诱导。研究结果有助于深入研究荒漠昆虫在低温下MAPK信号转导途径的作用机制。
英文摘要	[Aim] Mitogen activated protein kinase (MAPK) cascade is one of the important signal transduction pathways in cells. Ras GTPase-activating protein (RasGAP) gene RasGAP and c-Jun N-terminal kinase (JNK) gene JNK are an up-stream gene and a down-stream gene in MAPK pathway, respectively. This study aims to determine the expression profiles of RasGAP and JNK in the desert beetle Microdera punctipennis in response to low temperature. [Methods] A full length cDNA of RasGAP from M. punctipennis was cloned. The deduced amino acid sequence was analyzed, and the phylogynetic tree was constructed. The expression profiles of RasGAP and JNK in M. punctipennis exposed to low temperatures were detected by using real-time quantitative PCR. [Results] The ORF of RasGAP cDNA from M. punctipennis is 2 523 bp in length, and was named as MpRasGAP (GenBank accession no.: KM677930), encoding a polypeptide of 840 amino acids with the molecular weight of 96.594 kDa. The encoded protein MpRasGAP belongs to the RasGAP super family. Homology analysis showed that MpRasGAP shares 89% amino acid sequence identity with RasGAP from Tribolium castaneum. When M. punctipennis adults were exposed to 4℃ and -4℃ for 1 h, the mRNA levels of MpRasGAP were significantly upregulated as compared with that at room temperature (25℃). When M. punctipennis adults were exposed to 4℃ for 3 h or -4℃ for 1 h, the mRNA levels of MpJNK were also elevated significantly. [Conclusion] The results of this study demonstrate that the expression of both MpRasGAP and MpJNK in M. punctipennis can be induced by low temperature. Our results will help to further study the role of MAPK pathway in the desert insect under low temperature.
中文关键词	小胸鳖甲 ; 低温胁迫 ; Ras GTP酶激活蛋白 ; 基因克隆 ; mRNA水平 ; 实时定量PCR
英文关键词	Microdera punctipennis low temperature stress Ras GTPase-activating protein (RasGAP) gene cloning mRNA level real-time quantitative PCR
语种	中文
国家	中国
收录类别	CSCD
WOS类目	ENTOMOLOGY
WOS研究方向	Entomology
CSCD记录号	CSCD:5438936
来源机构	新疆大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/232960
作者单位	新疆大学生命科学与技术学院, 新疆生物资源基因工程重点实验室, 乌鲁木齐, 新疆 830046, 中国
推荐引用方式 GB/T 7714	阮梦鸽,李洁琼,孟闪闪,等. 荒漠昆虫小胸鳖甲Ras GTP酶激活蛋白基因MpRasGAP的克隆及低温表达分析[J]. 新疆大学,2015,58(4):367-374.
APA	阮梦鸽,李洁琼,孟闪闪,&马纪.(2015).荒漠昆虫小胸鳖甲Ras GTP酶激活蛋白基因MpRasGAP的克隆及低温表达分析.昆虫学报,58(4),367-374.
MLA	阮梦鸽,et al."荒漠昆虫小胸鳖甲Ras GTP酶激活蛋白基因MpRasGAP的克隆及低温表达分析".昆虫学报 58.4(2015):367-374.