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Rapid cloning of rearranged immunoglobulin genes from human hybridoma cells using mixed primers and the polymerase chain reaction

Larrick, James W; Danielsson, Lena; Brenner, Carol A; Abrahamson, Magnus LU ; Fry, Kirk E and Borrebaeck, Carl LU (1989) In Biochemical and Biophysical Research Communications 160(3). p.1250-1256
Abstract
A general method to directly obtain the DNA sequence of the variable regions of any immunoglobulin chain using a mixture of oligomer primers and the polymerase chain reaction (PCR) is described. Mixed oligonucleotide primers corresponding to the 5′ signal peptide and a conserved 3′ constant region primer were used for enzymatic amplification of each of the heavy and light chain variable regions of a human hybridoma producing a monoclonal antibody recognizing an epitope of gp120 of the human immunodeficiency virus 1. The amplified DNA segments were cloned and the sequence was determined for the heavy chain variable region. This method will greatly facilitate structural and functional studies of immunoglobulins by reducing the effort to... (More)
A general method to directly obtain the DNA sequence of the variable regions of any immunoglobulin chain using a mixture of oligomer primers and the polymerase chain reaction (PCR) is described. Mixed oligonucleotide primers corresponding to the 5′ signal peptide and a conserved 3′ constant region primer were used for enzymatic amplification of each of the heavy and light chain variable regions of a human hybridoma producing a monoclonal antibody recognizing an epitope of gp120 of the human immunodeficiency virus 1. The amplified DNA segments were cloned and the sequence was determined for the heavy chain variable region. This method will greatly facilitate structural and functional studies of immunoglobulins by reducing the effort to clone and sequence the members of the immunoglobulin as well as other multigene families. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemical and Biophysical Research Communications
volume
160
issue
3
pages
1250 - 1256
publisher
Elsevier
external identifiers
  • scopus:0024346547
ISSN
1090-2104
DOI
10.1016/S0006-291X(89)80138-X
language
English
LU publication?
yes
id
05530df1-ca51-431a-b0dc-60429ee13eb4 (old id 1104884)
date added to LUP
2008-08-07 08:24:46
date last changed
2017-08-20 04:28:14
@article{05530df1-ca51-431a-b0dc-60429ee13eb4,
  abstract     = {A general method to directly obtain the DNA sequence of the variable regions of any immunoglobulin chain using a mixture of oligomer primers and the polymerase chain reaction (PCR) is described. Mixed oligonucleotide primers corresponding to the 5′ signal peptide and a conserved 3′ constant region primer were used for enzymatic amplification of each of the heavy and light chain variable regions of a human hybridoma producing a monoclonal antibody recognizing an epitope of gp120 of the human immunodeficiency virus 1. The amplified DNA segments were cloned and the sequence was determined for the heavy chain variable region. This method will greatly facilitate structural and functional studies of immunoglobulins by reducing the effort to clone and sequence the members of the immunoglobulin as well as other multigene families.},
  author       = {Larrick, James W and Danielsson, Lena and Brenner, Carol A and Abrahamson, Magnus and Fry, Kirk E and Borrebaeck, Carl},
  issn         = {1090-2104},
  language     = {eng},
  number       = {3},
  pages        = {1250--1256},
  publisher    = {Elsevier},
  series       = {Biochemical and Biophysical Research Communications},
  title        = {Rapid cloning of rearranged immunoglobulin genes from human hybridoma cells using mixed primers and the polymerase chain reaction},
  url          = {http://dx.doi.org/10.1016/S0006-291X(89)80138-X},
  volume       = {160},
  year         = {1989},
}