Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids.
A hybrid cell line (R21/B1) containing a truncated human chromosome 6 (6pter-6q21) and a human Y chromosome on a hamster background was irradiated and fused to A23 (TK-) or W3GH (HPRT-) hamster cells. Clones containing expressed HLA class I genes (4/40) were selected using monoclonal antibodies. The...
Main Authors: | , , , , , , |
---|---|
Format: | Journal article |
Language: | English |
Published: |
1991
|
_version_ | 1797090966133276672 |
---|---|
author | Ragoussis, I Jones, T Sheer, D Shrimpton, A Goodfellow, P Trowsdale, J Ziegler, A |
author_facet | Ragoussis, I Jones, T Sheer, D Shrimpton, A Goodfellow, P Trowsdale, J Ziegler, A |
author_sort | Ragoussis, I |
collection | OXFORD |
description | A hybrid cell line (R21/B1) containing a truncated human chromosome 6 (6pter-6q21) and a human Y chromosome on a hamster background was irradiated and fused to A23 (TK-) or W3GH (HPRT-) hamster cells. Clones containing expressed HLA class I genes (4/40) were selected using monoclonal antibodies. These clones were recloned and analyzed with a panel of probes from the HLA region. One hybrid (4G6) contained the entire HLA complex. Two other hybrids (4J4 and 4H2) contained only the HLA class I region, while the fourth hybrid (5P9) contained HLA class I and III genes in addition to other genes located in the 6p21 chromosomal region. In situ hybridization showed that the hybrid cells contained more than one fragment of human DNA. Alu and LINE PCR products were derived from these cells and compared to each other as well as to products from two somatic cell hybrids having the 6p21 region in common. The PCR fragments were then screened on conventional Southern blots of the somatic cell hybrids to select a panel of novel probes encompassing the 6p21 region. In addition, the origin of the human DNA fragments in hybrid 4J4 was determined by regional mapping of PCR products. |
first_indexed | 2024-03-07T03:26:15Z |
format | Journal article |
id | oxford-uuid:b9227875-5938-4656-b843-b57788cdc594 |
institution | University of Oxford |
language | English |
last_indexed | 2024-03-07T03:26:15Z |
publishDate | 1991 |
record_format | dspace |
spelling | oxford-uuid:b9227875-5938-4656-b843-b57788cdc5942022-03-27T05:00:53ZIsolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b9227875-5938-4656-b843-b57788cdc594EnglishSymplectic Elements at Oxford1991Ragoussis, IJones, TSheer, DShrimpton, AGoodfellow, PTrowsdale, JZiegler, AA hybrid cell line (R21/B1) containing a truncated human chromosome 6 (6pter-6q21) and a human Y chromosome on a hamster background was irradiated and fused to A23 (TK-) or W3GH (HPRT-) hamster cells. Clones containing expressed HLA class I genes (4/40) were selected using monoclonal antibodies. These clones were recloned and analyzed with a panel of probes from the HLA region. One hybrid (4G6) contained the entire HLA complex. Two other hybrids (4J4 and 4H2) contained only the HLA class I region, while the fourth hybrid (5P9) contained HLA class I and III genes in addition to other genes located in the 6p21 chromosomal region. In situ hybridization showed that the hybrid cells contained more than one fragment of human DNA. Alu and LINE PCR products were derived from these cells and compared to each other as well as to products from two somatic cell hybrids having the 6p21 region in common. The PCR fragments were then screened on conventional Southern blots of the somatic cell hybrids to select a panel of novel probes encompassing the 6p21 region. In addition, the origin of the human DNA fragments in hybrid 4J4 was determined by regional mapping of PCR products. |
spellingShingle | Ragoussis, I Jones, T Sheer, D Shrimpton, A Goodfellow, P Trowsdale, J Ziegler, A Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids. |
title | Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids. |
title_full | Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids. |
title_fullStr | Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids. |
title_full_unstemmed | Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids. |
title_short | Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion gene transfer hybrids. |
title_sort | isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation fusion gene transfer hybrids |
work_keys_str_mv | AT ragoussisi isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids AT jonest isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids AT sheerd isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids AT shrimptona isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids AT goodfellowp isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids AT trowsdalej isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids AT zieglera isolationofprobesspecifictohumanchromosomalregion6p21fromimmunoselectedirradiationfusiongenetransferhybrids |