論文

査読有り
2002年6月

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

59
6
開始ページ
502
終了ページ
511
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1034/j.1399-0039.2002.590607.x
出版者・発行元
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.

Web of Science ® 被引用回数 : 35

リンク情報
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

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