家蚕微孢子虫极管蛋白1 (NbPTP1) 在果蝇S2细胞中的表达与糖基化修饰

Expression of Polar Tube Protein 1 (NbPTP1) from Nosema bombycis in Drosophila S2 cell lines and its glycosylation

摘要:

极管蛋白（Polar tube protein）是极管的主要成分，能特异性定位于微孢子虫极管，在微孢子虫侵染宿主过程中发挥重要作用。文中分析了家蚕微孢子虫极管蛋白1中潜在的O-、N-糖基化修饰位点，克隆了家蚕微孢子虫极管蛋白1全基因序列，并将其插入带有V5和His标签的真核表达载体pMT/Bip/V5-His A中，成功构建了pMT/Bip/V5-His A-NbPTP1重组质粒，经转染果蝇S2细胞后，发现NbPTP1基因能在果蝇细胞中高效表达。此外，Lectin blotting和β-消除反应分析结果表明：果蝇S2细胞内表达的NbPTP1具有O-糖基化修饰特征。以上结果为研究NbPTP1的糖基化修饰特征与其功能之间的关系提供了基础，有助于揭示微孢子虫侵染机制，建立可行有效的微孢子虫病诊断和防治措施。

Abstract:

The polar tube protein is the major component of polar tube, and can specifically locate on the polar tube of microsporidia and plays an important role in invasion host cell. In this study, we analyzed the potential O- and N- glycosylation sites in polar tube protein 1 from Nosema bombycis. NbPTP1 was successfully cloned to eukaryotic expression vector pMT/Bip/V5-His A, involved V5 and His tags. After transfection, NbPTP1 gene could be efficiently expressed in Drosophila S2 cells. In addition, Lectin blotting and beta elimination analysis showed that NbPTP1 expressed in Drosophila S2 cells was O-glycosylation. These studies provided a basis for understanding the relationship between glycosylation and function of NbPTP1, helped us to reveal the infection mechanism of microsporidia and established effective diagnosis and prevention methods for microsporidia.