Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells

Détails

ID Serval
serval:BIB_EEA160205082
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells
Périodique
PCR Methods and Applications
Auteur(s)
Berdoz  J., Monath  T. P., Kraehenbuhl  J. P.
ISSN
1054-9803 (Print)
Statut éditorial
Publié
Date de publication
04/1995
Volume
4
Numéro
5
Pages
256-64
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Apr
Résumé
We have designed a novel strategy for the isolation of the rearranged genomic fragments encoding the L-VH-D-JH and L-V kappa/lambda-J kappa/lambda regions of mouse immunoglobulin genes. This strategy is based on the PCR amplification of genomic DNA from mouse hybridomas using multiple specific primers chosen in the 5'-untranslated region and in the intron downstream of the rearranged JH/J kappa/lambda sequences. Variable regions with intact coding sequences, including full-length leader peptides (L) can be obtained without previous DNA sequencing. Our strategy is based on a genomic template that produces fragments that do not need to be adapted for recombinant antibody expression, thus facilitating the generation of chimeric and isotype-switched immunoglobulins.
Mots-clé
Amino Acid Sequence Animals Base Sequence Blotting, Northern Cell Line DNA Primers *Gene Rearrangement *Genes, Immunoglobulin Hybridomas/immunology Immunoglobulin Heavy Chains/genetics Immunoglobulin Variable Region/*genetics Immunoglobulin kappa-Chains/genetics Immunoglobulin lambda-Chains/genetics Mice Molecular Sequence Data Oligonucleotide Probes Polymerase Chain Reaction/*methods Pseudogenes
Pubmed
Web of science
Création de la notice
25/01/2008 16:05
Dernière modification de la notice
03/03/2018 22:33
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