Cloning and sequencing of human lambda immunoglobulin genes by the polymerase chain reaction.
Universal oligonucleotide primers, designed for amplifying and sequencing genes encoding the rearranged human lambda immunoglobulin variable region, were validated by amplification of the lambda light chain genes from four human heterohybridoma cell lines and in the generation of a cDNA library of human V lambda sequences from Epstein-Barr virus-transformed human peripheral blood lymphocytes. This technique allows rapid cloning and sequencing of human immunoglobulin genes, and has potential applications in the rescue of unstable human antibody-producing cell lines and in the production of human monoclonal antibodies.
Songsivilai
S
S
Molecular Immunopathology Unit, Medical Research Council Centre, Cambridge, GB.
Bye
J M
JM
Marks
J D
JD
Hughes-Jones
N C
NC
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005803:Genes, Immunoglobulin; D006801:Humans; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007146:Immunoglobulin lambda-Chains / Q000235:genetics*; D008214:Lymphocytes / Q000502:physiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping
D007135:Immunoglobulin Variable Region; D007146:Immunoglobulin lambda-Chains; D004247:DNA