黄曲条跳甲双孔道钾离子通道基因序列特征及不同组织的表达量分析

Characteristics and expression profile of two-P domain K^+ channels of Phyllotreta striolata(Fabricius)

昆虫细胞膜钾离子通道与多种生命活动密切相关。结合转录组测序及荧光定量PCR技术,鉴定和分析黄曲条跳甲(Phyllotreta striolata(Fabricius))钾离子通道基因的序列特征及不同组织器官的mRNA表达谱。结果表明:所获得的钾离子通道蛋白基因(twk)cDNA的开放阅读框为1 137 bp,共编码378个氨基酸残基。蛋白结构分析表明:其含有4个钾离子通道蛋白的跨膜区(M1～M4)和2个孔道结构域(1P～2P),分别位于M1与M2、M3与M4之间,即双孔道钾离子通道。序列保守性分析表明:1P的核心序列为甘氨酸-酪氨酸-甘氨酸(GYG)模体,2P的核心序列为甘氨酸-亮氨酸-甘氨酸(GLG)模体。荧光定量PCR分析表明,twk在黄曲条跳甲雌雄成虫的不同部位中都有表达,但是在精巢或卵巢、中肠和头部的表达量相对较高;雌、雄成虫各组织器官之间的相对表达量没有显著差异。

Potassium channel participate in the adjustment biology in a variety of physiological process.The expressed sequence tag（EST）of two-P domain K＋ channel（twk）was clustered and identified from transcriptome data of Phyllotreta striolata（Fabricius）.Gene expression profiles in different tissues were analyzed by quantitative real-time PCR.This cDNA was 1 137 bp in length with an open reading frame（ORF）encoding 378 amino acids.The putative protein molecular contained 4 transmembrane helixs（M1-M4）and 2 pore domains（1P-2P）,locating between M1 and M2,M3 and M4.Conservation elements analysis showed that the core sequences of 1P is GYG motif,and the core sequences of 2P is GLG motif.Quantitative real-time PCR assay showed that a relatively higher expression amount of twk gene was found in testis or ovary,midgut and head than that in other tissues.There were no significance differences of twk expression profile between male and female adults.