中国兽医科学2024，Vol.54Issue(4)：433-440,8.DOI:10.16656/j.issn.1673-4696.2024.0054
Rab18调控禽偏肺病毒C型复制的研究
Effect of Rab18 on the replication of avian metapneumovirus type C
摘要
Abstract
This work aimed to investigate how Rab18 influences the replication of avian metapneu-movirus type C(aMPV/C)in A549 cells.Confocal images were obtained by laser confocal microscope af-ter being infected with aMPV/C.The colocalization coefficient was analyzed by Image J software.The results showed that Rab18 and aMPV/C N protein were expressed in the cytoplasm and showed obvious co-localization,while no fluorescent signal of N protein was observed in the mock group.The co-local-ization coefficient between Rab18 and aMPV/C N protein was significantly higher than that in the mock group(P＜0.01).The plasmids(pCMV-Flag-Rab18 and pCMV-Flag)and nucleic acids(siRab18 and siNC)were transfected into A549 cells before being infected with aMPV/C,respectively.Cell samples and cell su-pernatant were collected for detection of the transcription level of aMPV/C Ngene,the expression lev-el of N protein,and viral titers by qPCR,Western-blot,and TCID50,respectively.The results showed that the transcription level of aMPV/C Ngene,the expression level of N protein,and the titer of virus were significantly increased after overexpression of Rab18,respectively(P＜0.01).On the contrary,the transcription level of aMPV/C Ngene,the expression level of N protein,and the viral titers were sig-nificantly decreased after knockdown expression of Rab18,respectively(P＜0.01).A549 cells were co-transfected with pCMV-mcherry-Rab18 and GFP-tagged aMPV/C genes before confocal imaging.The results showed that Rab18 could co-localize with aMPV/C proteins in the cytoplasm.HKE293T cells were co-trans-fected with pCMV-Flag-Rab18 and GFP-tagged aMPV/C genes before conducting the co-immunoprecipitation(co-IP)assay.The results showed that specific N,M2-1,and L2 protein bands were found in IP samples while other viral proteins were not detected.These results suggest that Rab18 influences aMPV/C replication via interaction with N,M2-1,and L2 proteins.The results provided a foundation for further analysis of the molecular mechanism of regulation aMPV/C replication by Rab18.关键词
禽偏肺病毒C型/Rab18/病毒复制/相互作用/表达水平Key words
avian metapneumovirus type C/Rab18/virus replication/interaction/expression level分类
农业科技引用本文复制引用
张正洲,王如嘉,亓宇翔,于瀚哲,孟闯,冯旭飞..Rab18调控禽偏肺病毒C型复制的研究[J].中国兽医科学,2024,54(4):433-440,8.基金项目
江苏省高等学校基础科研(自然科学)面上项目(21KJB230009) （自然科学）
江苏省人兽共患病学重点实验室开放课题项目(R2106) （R2106）
高等学校学科创新引智计划资助项目(D18007) （D18007）
江苏高校优势学科建设工程资助项目(PAPD) （PAPD）
