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Nucleic Acids Research, 1981, Vol. 9, No. 18 4485-4494
© 1981

MOLECULAR BIOLOGY

Complete sequence of an immunoglobulin mRNA using specific priming and the dideoxynucleotide method of RNA sequencing

Pamela H. Hamlyn, Michael J. Gait and Cesar Milstein

Laboratory of Molecular Biology The MRC Centre, Hills Road, Cambridge CB2 2QH, UK

Received June 30, 1981. The complete sequence of the mouse immunoglobulin kappa light chain MOPC 21 messenger RNA has been determined using a chain termination method and chemically synthesised deoxyoligonucleotides to initiate the synthesis of a DNA molecule complementary to the mRNA template. Five such oligonucleotide primers have been used for the sequence analysis of this messenger RNA. The approach is excellent for comparative studies of mouse {kappa}-chain mRNAs because they can be made on impure mRNA preparations.

The MOPC 21 light chain mRNA is 943 nucleotides in length excluding the poly (A) region. An unexpected finding was that there are only three bases in the 5' non-coding region and its significance in terms of ribosome binding is discussed; 87 code for the precursor or leader sequence of the protein, 642 for the mature protein and 211 for the 3' non-coding region. The codons for the precursor region allows the previously undetermined amino acid sequence to be predicted. In common with other precursor regions a high proportion of the predicted amino acids are hydrophobic.


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