Direct cDNA cloning of the rearranged immunoglobulin variable region.
A major problem in the study of multigene families is the effort required to clone and sequence these genes. We describe a method to rapidly clone and sequence immunoglobulin variable region gene sequences without constructing cDNA libraries. Because immunoglobulin variable-region genes are flanked by conserved sequences, we have been able to apply the polymerase chain reaction (PCR) to clone and sequence both the light- and heavy-chain rearranged immunoglobulin genes from small numbers of hybridoma cells. This method will greatly facilitate the construction of chimeric mouse/human monoclonal antibodies for immunoglobulin structural studies as well as for therapeutic use.
Chiang
Y L
YL
Genetic Therapy, Inc., Gaithersburg, MD 20878.
Sheng-Dong
R
R
Brow
M A
MA
Larrick
J W
JW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics; D001483:Base Sequence; D001709:Biotechnology; D002678:Chimera; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics*; D005784:Gene Amplification; D015321:Gene Rearrangement; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D004247:DNA