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

Author:
  • James W Larrick
  • Lena Danielsson
  • Carol A Brenner
  • Magnus Abrahamson
  • Kirk E Fry
  • Carl Borrebaeck
Publishing year: 1989
Language: English
Pages: 1250-1256
Publication/Series: Biochemical and Biophysical Research Communications
Volume: 160
Issue: 3
Document type: Journal article
Publisher: Elsevier

Abstract english

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.

Keywords

  • Biological Sciences

Other

Published
  • ISSN: 1090-2104
Carl B
Carl Borrebaeck
E-mail: carl.borrebaeck [at] immun.lth.se

Professor

Department of Immunotechnology

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