Messenger RNA from allotype‐defined rabbits directing the cell‐free synthesis of immunoglobulin heavy and light chains

摘要

AbstractIn vitrosynthesis studies were performed utilizing poly A(+)‐RNA and lymphocytes from the spleens of rabbits hyperimmunized withMicrococcus luteusorStreptococcus pneumoniae(type III). Poly A(+)‐RNA isolated after 4 M guanidinium thiocyanate extraction and oligo(dT)‐selection appeared to be undegraded on CH3HgOH‐agarose gel electrophoresis and demonstrated biological activity when translated in a rabbit reticulocyte cell‐free system. The electrophoretic patterns of the specifically immunoprecipitated cell‐free products were compared with those of Nonidet‐P40 extracts (lysates) and secretions (supernatants) from rabbit spleen lymphocyte cultures and serum proteins. Kappa light chains with specificballotypes, as well as immunoglobulin heavy chains, were identified. The efficient translation of mRNA of defined allotypes was a necessary prerequisite for production of characterized cDNA clones and identification of genomic sequences for rabbit immunoglobulin heavy and