Use of Mass spectrometry to identify Zika Virus NS1 interaction proteins
ZHANG Na1， WANG Hengrui2， MAO Yanna3， ZHANG Chenglin1， QIN Bin1， WANG Zhimin1
1.Sino-British Research Centre for Molecular Oncology， National Centre for International Research in Cell and Gene Therapy， School of Basic Medical Sciences， Academy of Medical Sciences， Zhengzhou University， Zhengzhou 450052， China；2.No 6 Middle School， Anyang， Henan 455000， China；3.Department of Hematology and Oncology， Children's Hospital Affiliated to Zhengzhou University， Zhengzhou 455000， China
Abstract：Zika Virus belongs to the genus Flavivirus in the Flaviviridae family. Although it has been discovered by humans for a long time， it has caused widespread concern until the massive outbreak of Brazil in South America in 2015. The Zika virus genome contain a single-stranded positive-stranded RNA， and its genome can encode and cleave three structural proteins which includes membrane glycoprotein， envelope protein and capsid protein and 7 non-structural proteins (NS1， NS2A， NS2B， NS3， NS4A， NS5). Here， we constructed and expressed NS1 fusion protein tagged with Flag and Strep in HEK293 cells， further we separated the protein bound to NS1 using immunoprecipitation assay. The binding proteins were analyzed and identified with high resolution mass spectrometry. Through the analysis of interacting proteins， 16 binding proteins shared by the two tags were found. Further pathway analysis confirmed that these proteins belong to multiple pathways related to viral transcription and viral replication. This research results provide important reference value for further research on Zika Virus replication mechanism and vaccine development.
2019, Vol. 53 
