Messenger RNA from allotype-defined rabbits directing the cell-free synthesis of immunoglobulin heavy and light chains.
In vitro synthesis studies were performed utilizing polyA(+)-RNA and lymphocytes from the spleens of rabbits hyperimmunized with Micrococcus luteus or Streptococcus pneumoniae (type III). PolyA(+)-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 specific b allotypes, 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 light chains.
D000818:Animals; D002474:Cell-Free System; D002478:Cells, Cultured; D011232:Chemical Precipitation; D004591:Electrophoresis, Polyacrylamide Gel; D007126:Immunoglobulin Allotypes / Q000235:genetics / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis*; D008214:Lymphocytes / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000276:immunology*; D011817:Rabbits; D012156:Reticulocytes / Q000032:analysis; D013154:Spleen / Q000032:analysis / Q000166:cytology
D007126:Immunoglobulin Allotypes; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D012333:RNA, Messenger