2002年6月
Efficient cloning and expression of HLA class I cDNA in human B-lymphoblastoid cell lines
TISSUE ANTIGENS
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- 巻
- 59
- 号
- 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.
- リンク情報
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- 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情報
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- DOI : 10.1034/j.1399-0039.2002.590607.x
- ISSN : 0001-2815
- ORCIDのPut Code : 47323580
- SCOPUS ID : 0036619417
- Web of Science ID : WOS:000178126000007