摘要
为了探究信号肽对A型流感病毒血凝素蛋白(hemagglutinin,HA)表达的影响,扩增了病毒的HA基因,利用双酶切方法将HA基因分别克隆至pCMV-Flag-N及pCMV-Flag-C载体中,成功获得了包含HA信号肽的SP-HA-Flag载体和缺失信号肽的Flag-HA载体,将重组质粒转染至293T细胞后利用Western blot及间接免疫荧光方法检测HA蛋白表达情况。结果表明,该信号肽显著下调HA蛋白的表达量,并调控HA在细胞内的定位:含信号肽序列的HA蛋白主要定位在细胞膜,而不含信号肽的HA蛋白则主要定位在细胞质。本研究为进一步研究流感病毒HA蛋白的生物学功能提供了基础信息。
To explore the effect of signal peptide on HA expression and localization in cells,we cloned the CDS region of the HA gene with or without signal peptide into the vector of pCMV-Flag-C and pCMV-Flag-N,respectively.The recombinant vectors were transfected into 293 T cells,meansured the expression of the HA protein by Western blot and indirect immunofluorescence assay.The results showed that the signal peptide reduced the expression level of HA.Moreover,the signal peptide had an effect on the localization of HA:HA with signal peptide located in the cell membrane,while HA without signal peptide located in the cytoplasm.This study provided basic information for further research on biological function of the HA protein of the influenza virus.
作者
郑屠园
李娅慧
周继勇
胡伯里
ZHENG Tuyuan;LI Yahui;ZHOU Jiyong;HU Boli(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;MOA Key Laboratory of Animal Virology/Department of Veterinary Medicine,Zhejiang University,Hangzhou 310058,China)
出处
《畜牧与兽医》
北大核心
2020年第8期64-67,共4页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31630077
31502084)。
关键词
血凝素
载体构建
信号肽
重组蛋白表达
HA
vector construction
signal peptide
recombinant protein expression
