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Estimation of the Number of Genes Specifying the Heavy Chain of Mouse Thymus Leukemia Antigens¹

Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, Illinois 60680

We have investigated the number of structural genes present in ASL-1 cells, a murine leukemia cell line which encodes the heavy chain of thymus leukemia (TL) antigens, a protein which is similar to the Class I histocompatibility antigens. TL-specific messenger RNA was purified from polysomes of ASL-1 cells by immunoprecipitation, and this messenger RNA translated in vitro to produce a M_r 42,000 protein which comigrated on acrylamide gel with nonglycosylated TL heavy chain. A ³²P-labeled comple-mentary DNA (cDNA) was synthesized by reverse transcription of the TL-specific messenger RNA as template. Analysis of reassociation kinetics of the ³²P-TL-cDNA with DNA from ASL-1 cells showed that the kinetics was indistinguishable from that obtained using a DNA encoding a single-copy gene (Cµ). An analysis was performed in which DNA from ASL-1 cells was subjected to digestion with each of three restriction enzymes and hybridized with ³²P-TL-cDNA according to the Southern "blot" technique. Two bands formed hybrids with the ³²P-TL-cDNA with each of three restriction enzymes used. These data are consistent with the presence of a small number of structural genes for the TL heavy chain in the genome of ASL-1 leukemia cells.

¹ Sponsored by USPHS Grant CA 27579-03.

² Present address: Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.

³ To whom requests for reprints should be addressed.

Received 1/18/83. Accepted 4/12/83.