title: [Liver tissue-specific stable expression of human CD81 molecule].
abstract: OBJECTIVE
  To express stably human CD81 gene on mouse hepatoma cell line Hepa 1-6 using liver specific promoter.
  
  
  METHODS
  RNA were isolated from human HepG2 cells which could be infected with hepatitis C virus. RT-PCR was carried out using human CD81 gene specific primers. Amplified fragments were cloned into pGEM-T vector. Albumin promoter and enhancer which were liver tissue specific were ligated to the 5'end of human CD81 gene and SV40 polyA sequence was fused with 3'end of CD81. The fused CD81 gene was inserted into eukaryotic expression vector pcDNA3 to construct a recombinant vector pcDNA3-Alb p-CD81 which was then transfected into Hepa 1-6 cells through lipofectamine mediation. Human CD81 mRNA transcription and its protein expression were detected by RT-PCR and FACS, respectively.
  
  
  RESULTS
  Sequence analysis showed that the cloned gene segment was human CD81 gene sequence. After transfection, transcripted human CD81 mRNA was obtained and human CD81 molecules were expressed stably on Hepa 1-6 cells.
  
  
  CONCLUSIONS
  The obtained positive cell clones which stably express HCV receptor human CD81 lay the foundation for further study on interactions between HCV envelope proteins and human CD81, screening of HCV infection blocking drugs and development of HCV infection mouse model.
authors.affiliation: Institute of Transfusion Medicine, Beijing 100850, China. jiasz@nic.bmi.ac.cn
authors.forename: Shuai-zheng
authors.initials: SZ
authors.lastname: Jia
authors.forename: Li-ping
authors.initials: LP
authors.lastname: Lu
authors.forename: Min-xia
authors.initials: MX
authors.lastname: Liu
authors.forename: Lin-sheng
authors.initials: LS
authors.lastname: Zhan
authors.forename: Hai-ping
authors.initials: HP
authors.lastname: Wang
authors.forename: Quan-li
authors.initials: QL
authors.lastname: Wang
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004195:Disease Models, Animal; D015870:Gene Expression; D006526:Hepatitis C / Q000209:etiology; D006801:Humans; D008099:Liver / Q000378:metabolism*; D051379:Mice; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D060225:Tetraspanin 28
chemical_list: D015703:Antigens, CD; C554798:CD81 protein, human; C554799:Cd81 protein, mouse; D018076:DNA, Complementary; D012333:RNA, Messenger; D060225:Tetraspanin 28