Position: Home > Articles > Cloning and sequence analysis of arginine kinase genes from two rice stem borer species
Plant Protection
2020
(3)
70-77
两种水稻蛀茎害虫精氨酸激酶基因的克隆及序列分析
作 者：
鲁艳辉;郑许松;田俊策;白琪;吕仲贤
单 位：
关键词：
水稻蛀茎害虫;稻蛀茎夜蛾;二化螟;AK基因;克隆;序列
摘 要：
精氨酸激酶(AK, EC 2.7.3.3)是昆虫体内唯一的磷酸原激酶,参与能量代谢。为探究AK基因的作用,本文利用本实验室建立的cDNA文库及RACE技术,分别从两种重要的水稻蛀茎害虫稻蛀茎夜蛾Sesamia inferens和二化螟Chilo suppressalis中克隆获得了AK基因的cDNA全长序列,并对其序列特征进行分析。AK基因分别命名为SinAK(稻蛀茎夜蛾,GenBank登录号:MK559476)和CsuAK(二化螟,MK559475),开放阅读框(ORF)长为1 065 bp,编码355个氨基酸,预测蛋白质分子量为40.0 kDa,等电点为5.88。氨基酸序列分析结果显示SinAK、CsuAK基因编码的氨基酸序列具有AK典型的功能位点保守序列:底物识别域S~(62)G~(63)V~(64)-Y~(67)和酶活性中心序列CP(S/T)N(I/L)GT。氨基酸序列比对及系统进化关系分析表明,SinAK与甜菜夜蛾、草地贪夜蛾和斜纹夜蛾的同源性最高,达95%以上,而CsuAK与印度谷螟、亚洲玉米螟的进化关系较近,同源性在95%左右。SinAK、CsuAK与其他昆虫AK的氨基酸序列同源性也高于80%,而与部分脊椎动物起同样作用的肌酸激酶(CK)的同源性低于40%,说明AK基因的功能是高度保守的,且与脊椎动物CK同源性很低。本研究结果为进一步研究SinAK和CsuAK的功能以及开发以AK基因为靶标的、对高等动物安全的害虫防治新技术奠定基础。
译 名：
Cloning and sequence analysis of arginine kinase genes from two rice stem borer species
作 者：
LU Yanhui;ZHENG Xusong;TIAN Junce;BAI Qi;Lü Zhongxian;State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection and Microbiology of Zhejiang Academy of Agricultural Sciences;
关键词：
rice stem borer;;Sesamia inferens;;Chilo suppressalis;;AK gene;;clone;;sequence
摘 要：
Arginine kinase(AK, EC 2.7.3.3) is an important regulation factor of energy metabolism in insect species. Aimed to explore the role of AK genes, in this study, two AK genes, named SinAK(GenBank accession number: MK559476), CsuAK(MK559475) were isolated from Sesamia inferens and Chilo suppressalis larvae, respectively, using cDNA library and RACE methods. Either cDNA sequence contained a 1 065 bp open reading frame(ORF) encoding a polypeptide of 355 amino acids with the predicted molecular weight and isoelectric point of 40.0 kD and 5.88, respectively. Amino acid sequence analysis showed that the SinAK and CsuAK sequences had the typical characteristics of arginine kinase, which contained the substrate recognition region S~(62)G~(63)V~(64)-Y~(67) and the active site CP(S/T)N(I/L)GT. The results of sequence comparison and phylogenetic analysis showed that SinAK shared more than 95% amino acid sequence identity with AKs from Spodoptera exigua, Spodoptera frugiperda and Spodoptera litura, while CsuAK shared more than 95% amino acid sequence identity with AKs from Plodia interpunctella and Ostrinia furnacalis. The amino acid sequence identities of SinAK and CsuAK with other insect AKs were also higher than 80%, while those with creatine kinases(CK) which play the same role in some vertebrates were lower than 40%. These results indicated that the function of AKs gene is highly conserved but they shared very low homology with vertebrate CK. Our results lay a foundation for further study of the functions of SinAK and CsuAK, and for the development of new pest control technologies targeting AK genes, which is safe for higher animal.
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