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Nucleic Acids Research, 1981, Vol. 9, No. 18 4547-4555
© 1981

MOLECULAR BIOLOGY

Purification and characterization of the messenger RNA for the heavy chain of rat immunoglobulin E

M.E. Zajdel-Blair*, G.E. Blair and H. Bennich

Department of Medical and Physiological Chemistry, The Biomedical Center Box 575, S-75123, Uppsala, Sweden Department of Biochemistry, University of Leicester Leicester LE1 7RH, UK

Received June 19, 1981.

The isolation and translational properties of rat inmiunoglobulin E (IgE) heavy chain mRNA are described. The mRNA has a sedimentation coefficient of approximately 18S, a chain length of about 2000 nucleotides and directs the synthesis in vitro of a polypeptide of 65000 molecular weight in an mRHA-dependent rabbit reticulocyte lysate. Inclusion of dog pancreatic microsomes in the cell-free translation system resulted in a heavy chain product of about 75000 molecular weight, presumably as a consequence of glycosylation in vitro. This species co-migrated in an SDS polyacrylamide gel with mature IgE heavy chain. Substantial purification of heavy chain mRNA was achieved by denaturing sucrose gradient centrifugation and agarose gel electrophoresis.

*Author to whom enquiries should be addressed at the Department of Biochemistry, University of Leicester, Leicester LE1 7RH, U.K.


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