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为探究禾谷缢管蚜Rhopalosiphum padi(Linnaeus)水通道蛋白基因Rp AQP1的序列特征及其在不同龄期的表达,利用RT-PCR和RACE技术克隆了Rp AQP1基因的c DNA全序列,采用生物信息学软件分析了Rp AQP1编码蛋白的特性,利用qRT-PCR分析了Rp AQP1在不同龄期的表达量。结果显示:禾谷缢管蚜水通道蛋白基因Rp AQP1的c DNA全长为1 216 bp,其750 bp的开放阅读框编码250个氨基酸,蛋白分子量为27.36 k D。Rp AQP1属于水通道蛋白亚家族DRIP(果蝇内嵌蛋白Drosophila integral protein)的一员,具有6个跨膜区,2个保守的NPA结构单元和1个压缩区域Ar/R;qRT-PCR结果显示,Rp AQP1在整个发育历期均有表达,其在2龄若蚜中表达水平最高,是成蚜表达量的1.432倍;在4龄若蚜中表达量最低,为成蚜表达量的0.444倍,显著低于其它龄期,而其余各龄期Rp AQP1表达量无显著差异。
In order to explore the sequence characteristics of Rhpalosiphum padi (Linnaeus) aquaporin gene Rp AQP1 and its expression at different stages, the full-length cDNA of Rp AQP1 gene was cloned by RT-PCR and RACE techniques. Informatics software analyzed the characteristics of Rp AQP1 encoding protein and analyzed the expression level of Rp AQP1 at different stages by qRT-PCR. The results showed that the full length cDNA of Rp AQP1 was 1 216 bp. The 750 bp open reading frame (ORF) encoded 250 amino acids with a molecular weight of 27.36 kD. Rp AQP1 is a member of the aquaporin subfamily DRIP (Drosophila integral protein), with 6 transmembrane regions, 2 conserved NPA structural units and 1 compressed region Ar / R; qRT-PCR results The results showed that Rp AQP1 was expressed throughout the entire developmental period and was the highest in the 2nd instar nymphs, which was 1.432 times of that of the adult aphids. The expression level of Rp AQP1 was the lowest in the 4th instar nymphs, which was 0.444 Fold, significantly lower than other age groups, while the rest of the age Rp AQP1 expression was no significant difference.