title: [Construction of recombinant vectors expressing antisense RNA to CCR5 and expression in eukaryotic cells].
abstract: BACKGROUND
  To construct recombinant vector expressing antisense RNA to CCR5 in eukaryotic cells and obtain recombinant pseudovirus, which will be used to block HIV-1 infection.
  
  
  METHODS
  The DNA fragment targeted against the initional part of CCR  5 mRNA translation was amplified by using RT-PCR from peripheral blood mononuclear cells (PBMCs) and cloned into retroviral vector pLXSN, then transfected into packaging cell (PA317) with lipofectAMINE. After 2-3 weeks selecting with G418, the pseudovirion in the survival cell's supernatant was detected with RT-PCR (FQ),then was used to infect NIH/3T3 cell.
  
  
  RESULTS
  The psuedovirion packed from expression vector of sense/antisense RNA to CCR5 had infected NIH/3T3 cell successfully. The vector had incorporated into its genome and transcripted into RNA.
  
  
  CONCLUSIONS
  The gene fragment of antisense RNA to CCR5 could be obtained from PBMCs and transfected into eukaryotic cell with retroviral vector. The results made a great foundation for studying its inhibiting effect on HIV-1 infection.
authors: Department of Infectious Disease, Peking University First Hospital, Beijing 100034, China.
authors: Huichun
authors: H
authors: Xing
authors: Xiaoyuan
authors: X
authors: Xu
authors: Qinhuan
authors: Q
authors: Wang
authors: Min
authors: M
authors: Yu
authors: Weibo
authors: W
authors: Gong
authors: Yiming
authors: Y
authors: Shao
mesh_terms: D016475:3T3 Cells; D000818:Animals; D005057:Eukaryotic Cells / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D051379:Mice; D010957:Plasmids / Q000235:genetics; D016372:RNA, Antisense / Q000235:genetics*; D019713:Receptors, CCR5 / Q000235:genetics*; D014162:Transfection
chemical_list: D016372:RNA, Antisense; D019713:Receptors, CCR5