摘要
蛋白激酶C受体l(Receptor for Activated C Kinase 1,RACKl)是WD40家族的成员,其作为稳定蛋白激酶C(Protein Kinases C,PKC)的受体,参与了多种生理活动。课题组前期从转录组数据中筛选获得了草地贪夜蛾(Spodoptera frugiperda)的RACK1序列,该研究通过生物信息软件对SfRACK1进行生物信息学分析,并以氨基酸序列为基础与其他昆虫的RACK1进行同源性比较;同时分析RACK1在草地贪夜蛾不同发育阶段不同组织以及NPV刺激后不同时间段的表达模式,探究草地贪夜蛾RACK1的功能以及该基因是否应答NPV刺激。结果表明:草地贪夜蛾RACK1的ORF序列长度为960 bp,可编码319个氨基酸,包含有7个重复的WD40结构域,预测其分子量和等电点分别为35.9 kDa和8.07,未发现信号肽和跨膜结构;多重序列比对及进化树分析表明,SfRACK1与棉铃虫、小菜蛾等鳞翅目昆虫亲缘关系较近;组织表达及时空表达结果表明,SfRACK1基因在脂肪体及成虫期具有较高的表达水平,而在表皮和卵中的表达量较低;当草地贪夜蛾受到NPV侵染后,其Sf RACK1基因在中肠的相对表达水平显著高于对照组。以上研究结果表明,SfRACK1基因在草地贪夜蛾正常生长发育过程中发挥了重要作用,而该基因可以响应NPV的侵染。
Receptor for Activated C Kinase 1(RACKl)is a member of the WD40 family.As a receptor for stabilizing Protein Kinase C(PKC),it is involved in a variety of physiological activities.In this study,RACK1 sequence of Spodoptera frugiperda was screened from transcriptome data,and its complete open reading frame(ORF)sequence was cloned by PCR technology.The bioinformatics analysis of Spodoptera frugiperda RACK1(SfRACK1)was carried out by bioinformatics software,which found that its ORF sequence was 960 bp in length,encoding 319 amino acids;SfRACK1 were predicted to be a 35.9 kDa relative molecular mass and 8.07isoelectric poin,containing 7 repeated WD40 domains;additionally,the SfRACK1 had no signal peptide and transmembrane structure.Multiple sequence alignment and phylogenetic tree analysis showed that SfRACK1 was closely related to Lepidopteran insects such as Helicoverpa armigera and Plutella xylostell.The RT-qPCR analysis results of tissue expression and spatio-temporal expression showed that the Sf RACK1 had a higher expression level in the fat body and adult stage,but a lower expression level in the integument and eggs.Following AcMNPV infection,the relative expression level of SfRACK1 in the midgut of the experimental group was significantly higher than that of the control group(fed on ddH2O).These findings indicate that SfRACK1 gene plays an important role in the normal growth and development of S.frugiperda,and it can respond to NPV infection.
作者
何凯
邓凡
严哲钰
张尚志
HE Kai;DENG Fan;YAN Zhe-yu;ZHANG Shang-zhi(College of Agriculture and Biotechnology,Hunan University of Humanities,Science and Technology,Loudi 417000,PRC;Hunan Provincial Collaborative Innovation Center for Field Weeds Control,Loudi 417000,PRC)
出处
《湖南农业科学》
2023年第1期1-8,共8页
Hunan Agricultural Sciences
基金
湖南人文科技学院大学生创新创业训练计划(S202110553018)
湖南省自然科学青年基金(2022JJ40195)
湖南省教育厅科学研究重点项目(21A0560)。
