UMMS Affiliation

Department of Molecular Genetics and Microbiology

Publication Date

2000-04-17

Document Type

Article

Subjects

Animals; B-Lymphocytes; Base Sequence; Cell Line; DNA; DNA Primers; Escherichia coli; Immunoglobulin Isotypes; *Immunoglobulin Switch Region; Mice; Mice, Inbred BALB C; Mice, Nude; Molecular Sequence Data; Plasmids; Polymerase Chain Reaction; Transcription, Genetic; Transfection; Transformation, Genetic

Disciplines

Life Sciences | Medicine and Health Sciences | Women's Studies

Abstract

Immunoglobulin class switch recombination (SR) occurs by a B cell-specific, intrachromosomal deletional process between switch regions. We have developed a plasmid-based transient transfection assay for SR to test for the presence of transacting switch activities. The plasmids are novel in that they lack a eukaryotic origin of DNA replication. The recombination activity of these switch substrates is restricted to a subset of B cell lines that support isotype switching on their endogenous loci and to mitogen-activated normal splenic B cells. The factors required for extrachromosomal plasmid recombination are constitutively expressed in proliferating splenic B cells and in B cell lines capable of inducibly undergoing immunoglobulin SR on their chromosomal genes. These studies suggest that mitogens that induce switching on the chromosome induce accessibility rather than switch recombinase activity. Finally, we provide evidence for two distinct switching activities which independently mediate mu-->alpha and mu-->gamma3 SR.

Keywords

immunoglobulin, isotype switch, plasmid assay, transient transfection, PCR

Rights and Permissions

Publisher PDF posted as allowed by the publisher's terms of use policy at: http://www.rupress.org/terms. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof.

DOI of Published Version

10.1084/jem.191.8.1365

Source

J Exp Med. 2000 Apr 17;191(8):1365-80.

Journal/Book/Conference Title

The Journal of experimental medicine

Related Resources

Link to article in PubMed

PubMed ID

10770803

Creative Commons License

Creative Commons Attribution-Noncommercial 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 License.

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