title: [Expression and assembly of rotavirus-like particles in insect cells mediated by recombinant Bombyx mori MultiBac].
abstract: OBJECTIVE
  To construct recombinant baculoviruses co-expressing three structural genes vp2, vp6 and vp7 of rotavirus, and assemble rotavirus-like particles (VLPs) in BmN cells.
  
  
  METHODS
  Human group A rotavirus was cultivated in MA104 cells, and the RNA was extracted and the three genes were obtained by RT-PCR. The PCR products were inserted into the transfer vectors pFBDM and pUCDM, respectively. A enhanced green fluorescent protein gene (egfp) driven by IE1 promoter was introduced into pFBDM to investigate the efficiency of infection. The expression baculoviruse was constructed by Tn7 and Cre-LoxP recombinant and transfected into BmN cells. The gene expression was determined by detecting 6-His tag fused into VP7 C-terminus, and the assembled VLPs were observed by transmission electron micrography.
  
  
  RESULTS
  Three genes of rotavirus were cloned and BmMultiBac was constructed. The genes were expressed and the rotavirus-like particles assembled in BmN cells successfully as verified by ELISA and electron microscope.
  
  
  CONCLUSIONS
  We have successfully constructed the recombinant baculovirus co-expressing the 3 structural genes of rotavirus, which provide the basis for producing protein complex containing multiple subunits and investigation of the structure of the macromolecules.
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou 510642, China. maclong@yahoo.cn
authors.forename: Hu
authors.initials: H
authors.lastname: Long
authors.forename: Lun-guang
authors.initials: LG
authors.lastname: Yao
authors.forename: Shan-shan
authors.initials: SS
authors.lastname: Wang
authors.forename: Shin-xin
authors.initials: SX
authors.lastname: Chen
authors.forename: Pei-chan
authors.initials: PC
authors.lastname: Tan
authors.forename: Jing-chen
authors.initials: JC
authors.lastname: Sun
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D012831:Bombyx / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics*; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D012401:Rotavirus / Q000235:genetics* / Q000378:metabolism
chemical_list: D036022:Capsid Proteins