Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells
Details
Serval ID
serval:BIB_EEA160205082
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells
Journal
PCR Methods and Applications
ISSN
1054-9803 (Print)
Publication state
Published
Issued date
04/1995
Volume
4
Number
5
Pages
256-64
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Apr
Research Support, Non-U.S. Gov't --- Old month value: Apr
Abstract
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.
Keywords
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
Create date
25/01/2008 16:05
Last modification date
20/08/2019 17:16