Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica
Two hundred thirty-seven independent somatic cell hybrids have been obtained between opossum (Monodelphis domestica) splenocytes, bone marrow cells, or primary fibroblasts, and HPRT-deficient or TK-deficient Chinese hamster, mouse, American mink, or common vole fibroblast lines. Because extreme segr...
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crskarger:10.1159/000134528 2024-10-13T14:06:43+00:00 Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica Nesterova, T.B. Isaenko, A.A. Matveeva, N.M. Shilov, A.G. Rubtsov, N.B. Vorobieva, N.V. Rubtsova, N.V. VandeBerg, J.L Zakian, S.M. 1997 http://dx.doi.org/10.1159/000134528 https://www.karger.com/Article/Pdf/134528 en eng S. Karger AG https://www.karger.com/Services/SiteLicenses https://www.karger.com/Services/SiteLicenses Cytogenetic and Genome Research volume 76, issue 3-4, page 115-122 ISSN 1424-8581 1424-859X journal-article 1997 crskarger https://doi.org/10.1159/000134528 2024-09-18T04:06:50Z Two hundred thirty-seven independent somatic cell hybrids have been obtained between opossum (Monodelphis domestica) splenocytes, bone marrow cells, or primary fibroblasts, and HPRT-deficient or TK-deficient Chinese hamster, mouse, American mink, or common vole fibroblast lines. Because extreme segregation and fragmentation of marsupial chromosomes commonly occurs in eutherian × marsupial somatic cell hybrids, we developed a rapid primary screening method that enables the identification of primary clones containing a large amount of opossum DNA 20–25 d after fusion. This method, which depends on in situ hybridization of biotin-labeled total opossum DNA on interphase nuclei of hybrid cells fixed on the bottom of microwell plates, was used to screen the 237 hybrid clones; 52 of them had a substantial amount of opossum DNA. G-banding and in situ hybridization of biotin labeled total opossum DNA on metaphase spreads of the clones enabled identification of 17 hybrid clones containing from two to seven intact chromosomes of M. domestica on the background of Chinese hamster or vole chromosomes. The hybrid clones with intact opossum chromosomes are used in a panel constructed for mapping the opossum genome. Initial mapping results from these clones have led to the tentative assignment of GPI and GOT1 to chromosome 1; 6PGD to chromosome 4; LDHA to chromosome 5; LDHB to chromosome 8; and PGK and G6PD to the X chromosome. On the basis of indirect evidence we also tentatively assigned HPRT to the X chromosome and TK to chromosome 5 of M. domestica. These are the first tentative chromosomal assignments by any technique for this species. Article in Journal/Newspaper Common vole Karger Cytogenetic and Genome Research 76 3-4 115 122 |
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English |
description |
Two hundred thirty-seven independent somatic cell hybrids have been obtained between opossum (Monodelphis domestica) splenocytes, bone marrow cells, or primary fibroblasts, and HPRT-deficient or TK-deficient Chinese hamster, mouse, American mink, or common vole fibroblast lines. Because extreme segregation and fragmentation of marsupial chromosomes commonly occurs in eutherian × marsupial somatic cell hybrids, we developed a rapid primary screening method that enables the identification of primary clones containing a large amount of opossum DNA 20–25 d after fusion. This method, which depends on in situ hybridization of biotin-labeled total opossum DNA on interphase nuclei of hybrid cells fixed on the bottom of microwell plates, was used to screen the 237 hybrid clones; 52 of them had a substantial amount of opossum DNA. G-banding and in situ hybridization of biotin labeled total opossum DNA on metaphase spreads of the clones enabled identification of 17 hybrid clones containing from two to seven intact chromosomes of M. domestica on the background of Chinese hamster or vole chromosomes. The hybrid clones with intact opossum chromosomes are used in a panel constructed for mapping the opossum genome. Initial mapping results from these clones have led to the tentative assignment of GPI and GOT1 to chromosome 1; 6PGD to chromosome 4; LDHA to chromosome 5; LDHB to chromosome 8; and PGK and G6PD to the X chromosome. On the basis of indirect evidence we also tentatively assigned HPRT to the X chromosome and TK to chromosome 5 of M. domestica. These are the first tentative chromosomal assignments by any technique for this species. |
format |
Article in Journal/Newspaper |
author |
Nesterova, T.B. Isaenko, A.A. Matveeva, N.M. Shilov, A.G. Rubtsov, N.B. Vorobieva, N.V. Rubtsova, N.V. VandeBerg, J.L Zakian, S.M. |
spellingShingle |
Nesterova, T.B. Isaenko, A.A. Matveeva, N.M. Shilov, A.G. Rubtsov, N.B. Vorobieva, N.V. Rubtsova, N.V. VandeBerg, J.L Zakian, S.M. Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica |
author_facet |
Nesterova, T.B. Isaenko, A.A. Matveeva, N.M. Shilov, A.G. Rubtsov, N.B. Vorobieva, N.V. Rubtsova, N.V. VandeBerg, J.L Zakian, S.M. |
author_sort |
Nesterova, T.B. |
title |
Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica |
title_short |
Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica |
title_full |
Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica |
title_fullStr |
Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica |
title_full_unstemmed |
Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica |
title_sort |
novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of monodelphis domestica |
publisher |
S. Karger AG |
publishDate |
1997 |
url |
http://dx.doi.org/10.1159/000134528 https://www.karger.com/Article/Pdf/134528 |
genre |
Common vole |
genre_facet |
Common vole |
op_source |
Cytogenetic and Genome Research volume 76, issue 3-4, page 115-122 ISSN 1424-8581 1424-859X |
op_rights |
https://www.karger.com/Services/SiteLicenses https://www.karger.com/Services/SiteLicenses |
op_doi |
https://doi.org/10.1159/000134528 |
container_title |
Cytogenetic and Genome Research |
container_volume |
76 |
container_issue |
3-4 |
container_start_page |
115 |
op_container_end_page |
122 |
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1812812927458082816 |