title: [Construction of a eukaryotic expression vector harboring the small interfering RNA targeting HCMV-IE1 gene and its gene silencing efficiency].
abstract: OBJECTIVE
  To investigate the effect of RNA interference targeting human cytomegalovirus immediate early gene 1 (HCMV- IE1) on the gene expression in vitro.
  
  
  METHODS
  According to the sequence of HCMV-IE1 gene, the small interfering RNA (siRNA) sequences were designed and introduced into the eukaryotic expression vector containing the U6 promoter. After verification by sequence analysis, the recombinant eukaryotic plasmid (pHCMV-IE1i) was transfected into HEL HCMVAD169 cells. The effectiveness of HCMV-IE1 gene silencing was investigated by fluorescence microscopy, flow cytometry and RT-PCR.
  
  
  RESULTS
  Sequence analysis confirmed successful construction of the recombinant eukaryotic expression plasmid pHCMV-IE1i. The expression of HCMV-IE1 was effectively suppressed by pHCMV-IE1i transfection in HEL cells as shown by fluorescence microscopy, flow cytometry (P < 0.05) and RT-PCR (P < 0.05).
  
  
  CONCLUSIONS
  The expression of HCMV-IE1 can be effectively suppressed by RNA interference technique in vitro, which provides experimental data for prevention and treatment of HCMV infection.
authors.affiliation: First Affiliated Hospital of Shenzhen University/ Second Shenzhen People's Hospital, Shenzhen 518035, China. gracexn2001@hotmail.com
authors.forename: Ni
authors.initials: N
authors.lastname: Xie
authors.forename: Yan-ping
authors.initials: YP
authors.lastname: Han
authors.forename: Jin-bin
authors.initials: JB
authors.lastname: Wu
authors.forename: Jian-hui
authors.initials: JH
authors.lastname: Yuan
mesh_terms: D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D017781:Genes, Immediate-Early; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins / Q000096:biosynthesis* / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D017874:Immediate-Early Proteins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C044582:immediate-early proteins, cytomegalovirus