龟纹瓢虫胰岛素信号通路AKT、MAPK、RPS6KB基因序列结构与时空表达模式分析

Sequence Structure and Spatiotemporal Expression Analysis of AKT,MAPK and RPS6KB Genes of Insulin Signaling Pathway in Propylea japonica

为明确龟纹瓢虫Propylea japonica胰岛素信号通路中AKT、MAPK和RPS6KB三个基因的序列结构和时空表达模式,基于龟纹瓢虫全基因组序列数据,选取AKT、MAPK和RPS6KB三个基因全长进行序列结构分析,蛋白序列结构分析发现其分别编码522、332和375个氨基酸。利用qRT-PCR检测其在龟纹瓢虫雌虫不同发育时期(羽化后1、3、5、7 d)、不同组织(头、胸、腹、肠)中的表达模式,结果显示AKT、MAPK和RPS6KB基因表达量均在龟纹瓢虫雌虫羽化第7 d显著上调,并达到最高水平;AKT在雌虫羽化第1 d和3 d高表达部位均为头部,第5 d和7 d高表达部位分别为肠道和胸部;MAPK基因在龟纹瓢虫雌虫羽化第1 d、3 d、5 d和7 d,头部表达水平均显著高于胸、腹和肠道;RPS6KB基因在其羽化后第1 d、3d、5d和7d,腹部表达水平均显著高于头、胸和肠道。AKT、MAPK、RPS6KB基因在龟纹瓢虫不同发育时期及不同组织中的表达量存在差异,为进一步研究胰岛素信号通路在龟纹瓢虫生殖发育中的作用奠定了基础。

To determine the sequence structure and spatiotemporal expression patterns of AKT,MAPK and RPS6KB genes in the insulin signaling pathway of Propylea japonica,the full-length genes of AKT,MAPK and RPS6KB were acquired from P.japonica genome.Protein sequence structure analysis showed that the genes encode 522,332,and 375 amino acids,respectively.Expression profiles of the genes in different developmental stages(1,3,5,7 days after eclosion)and tissues(head,thorax,abdomen,intestine)of P.japonica female adults were investigated through the method of real-time quantitative PCR.The results showed that AKT,MAPK and RPS6KB were significantly up-regulated in the 7-d old females and reached the highest level.AKT was highly expressed in the head in the 1-d and 3-d old females,and highly expressed in the intestine and thorax in the 5-d and 7-d old females,respectively.The expression level of MAPK gene in the head was significantly higher than that in the thorax,abdomen and intestine in the 1-d,3-d,5-d and 7-d old females.Expression level of RPS6KB gene was the highest in abdomen in the1-d,3-d,5-d and 7-d old females,and significantly higher than that in head,thorax and intestine.Taken together,the expression levels of AKT,MAPK and RPS6KB genes differ in different developmental stages and tissues of P.japonica,which serves as a foundation for further study on the role of insulin signaling pathway in reproductive development of P.japonica..