Papers

Peer-reviewed
Jun, 2002

Efficient cloning and expression of HLA class I cDNA in human B-lymphoblastoid cell lines

TISSUE ANTIGENS
  • Y Akatsuka
  • ,
  • TA Goldberg
  • ,
  • E Kondo
  • ,
  • EG Martin
  • ,
  • Y Obata
  • ,
  • Y Morishima
  • ,
  • T Takahashi
  • ,
  • JA Hansen

Volume
59
Number
6
First page
502
Last page
511
Language
English
Publishing type
Research paper (scientific journal)
DOI
10.1034/j.1399-0039.2002.590607.x
Publisher
BLACKWELL MUNKSGAARD

Analysis of HLA restriction specificity is one of the important steps in characterizing T cell clones, This usually requires, either it panel of HLA-typed cells or HLA cDNA transfectants. Although preparation of HLA cDNA transfectants is laborious, utilization of transfectants is advantageous when a suitable panel is not available due to linkage disequilibrium or rarity of the HLA allele of interest. In this report, we describe an efficient and rapid HLA cloning and expression system. Three sets of PCR primers specific for HLA-A, B and C loci were designed by extensively sequencing 5'- and 3'- untranslated regions of HLA class 1 genes. The PCR-amplified products were introduced into modified Phoenix retrovirus vectors containing a puromycin resistant gene under the control of a LTR promotor. Gibbon ape leukemia virus (GALV)-pseudotyped retrovirus was produced and infected into B-lymphoid cell lines, Following expansion in selection media, more than 80% of cells expressed transduced HLA at a comparable level to that normally expressed. These results indicate that locus-specific PCR cloning and utilization of GALV-pseudotyped retroviral vector can be an effective and relatively efficient tool for constructing a panel of different HLA transfectants.

Link information
DOI
https://doi.org/10.1034/j.1399-0039.2002.590607.x
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000178126000007&DestApp=WOS_CPL
URL
http://www.scopus.com/inward/record.url?eid=2-s2.0-0036619417&partnerID=MN8TOARS
URL
http://orcid.org/0000-0003-0995-9405
ID information
  • DOI : 10.1034/j.1399-0039.2002.590607.x
  • ISSN : 0001-2815
  • ORCID - Put Code : 47323580
  • SCOPUS ID : 0036619417
  • Web of Science ID : WOS:000178126000007

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