title: [Construction and expression of idiotype TCR Valpha1-pIRES-TCR Vbeta8 eukaryotic vectors in vitro].
abstract: OBJECTIVE
  To construct the eukaryotic vectors with idiotype TCR Valpha1-pIRES-TCR Vbeta8 of Jurkat cell line and investigate their expression in vitro after transferred into eukaryotic cells.
  
  
  METHODS
  TCR Valpha and Vbeta subfamilies of Jurkat cells were analyzed by using RT-PCR and genescan technique. Then the monoclonal TCR Valpha1 and Vbeta8 genes of Jurkat cells were cloned into multiple clone site (MCS) A and MCS B of the eukaryotic vector pIRES respectively. Its sequences were identified by restriction enzyme cutting and sequence analysis. The expression of TCR mRNA and idiotypic protein in transferred A549 and Molt4 cells was tested by RT-PCR, indirect immunophenotyping fluorescein dyeing and flow cytometry, respectively.
  
  
  RESULTS
  Two recombinavot eukaryotic vectors with idiotype TCR Valpha1-pIRES-TCR Vbeta8 were developed successfully and the expression of both idiotypic TCR mRNA and protein was detected in transferred A549 and Molt4 cells.
  
  
  CONCLUSIONS
  The successful construction of the two eukaryotic vectors with idiotype TCR Valpha1-pIRES-TCR Vbeta8 will provide a basic method for further study of the specific TCR gene modified T cells.
authors.affiliation: Institute of Hematology, Medical College of Jinan University, Guangzhou 510632, China. stlichuang@163.com
authors.forename: Chuang
authors.initials: C
authors.lastname: Li
authors.forename: Xiu-li
authors.initials: XL
authors.lastname: Wu
authors.forename: Yang-qiu
authors.initials: YQ
authors.lastname: Li
authors.forename: Yu-bing
authors.initials: YB
authors.lastname: Zhou
authors.forename: Shao-hua
authors.initials: SH
authors.lastname: Chen
authors.forename: Li-jian
authors.initials: LJ
authors.lastname: Yang
authors.forename: Kang-er
authors.initials: KE
authors.lastname: Zhu
mesh_terms: D056890:Eukaryota / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D011948:Receptors, Antigen, T-Cell