Direct cDNA cloning of the rearranged immunoglobulin variable region |
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Authors: | Y L Chiang R Sheng-Dong M A Brow J W Larrick |
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Institution: | Genetic Therapy, Inc., Gaithersburg, MD 20878. |
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Abstract: | 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. |
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