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PMID: 3034595 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Immunoglobulin heavy chain switch region recombination within a retroviral vector in murine pre-B cells.

The EMBO journal ·Vol. 6 ·No. 3 ·1987-03-00 ·Pages 577-84

Ott DE, Alt FW, Marcu KB

Abstract

We have employed a retroviral vector, ZN(Smu/S gamma 2b)tk1, as a substrate for detecting the presence of immunoglobulin heavy chain constant region (CH) gene switch (S) recombination activity in murine pre-B cells. ZN(Smu/S gamma 2b)tk1 contains a neomycin (neo) resistance gene in addition to the herpes simplex virus thymidine kinase (Htk) gene which is positioned between murine Smu and S gamma 2b sequences. Stable acquisition of the ZN(Smu/S gamma 2b)tk1 vector was selected in G-418 and switch region recombination within these proviruses was selected by resistance to the drug bromodeoxyuridine (BUdR). Fluctuation analyses of ZN(Smu/S gamma 2b)tk1 infected 18-8tk- and 38B9tk- pre-B lines revealed Htk gene inactivations with apparent frequencies of 5 X 10(-5) and 1 X 10(-5) events/cell/generation, respectively, while G-418 resistant Ltk- fibroblasts lost the HTK phenotype at an apparent rate of 4 X 10(-8). Southern blot analysis demonstrated that switch recombination caused the deletion of the Htk gene in all pre-B clones examined while the loss of Htk in Ltk- clones was not mediated by S region recombination. In 21 out of 24 pre-B clones, the recombinations involved the tandemly repetitive portions of the Smu and S gamma 2b sequences. These results demonstrate that the CH gene S region segments inserted into ZN(Smu/S gamma 2b)tk1 are sufficient for B-cell-specific recombination/deletion within the S region tandem repeats.

MeSH Terms
Animals B-Lymphocytes/immunology Bromodeoxyuridine/pharmacology Cell Line Drug Resistance Genes Genes, Viral Genetic Vectors Immunoglobulin Heavy Chains/genetics Immunoglobulin gamma-Chains/genetics Immunoglobulin mu-Chains/genetics Mice Recombination, Genetic Simplexvirus/enzymology,genetics Thymidine Kinase/genetics
Chemicals
Immunoglobulin Heavy Chains Immunoglobulin gamma-Chains Immunoglobulin mu-Chains Thymidine Kinase Bromodeoxyuridine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ott D E
Alt F W
Marcu K B
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1987-03-00
Pages
577-84
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553436
Subset
IM
Grants
NIAID NIH HHS · AI-20047 · United States
NIAID NIH HHS · AI00426 · United States
NIGMS NIH HHS · GM26939 · United States
External Links
PubMed source PMC Full Text View journal details
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