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Nucleic Acids Research, 1982, Vol. 10, No. 5 1495-1502
© 1982

MOLECULAR BIOLOGY

Chicken histone H5: selection of a cDNA recombinant using an extended synthetic primer

P.A. Krieg*, A.J. Robins*, M.J. Gait{dagger}, R.C. Titmas{dagger} and J.R.E. Wells*

*Department of Biochemistry, University of Adelaide South Australia 5001, UK {dagger}MRC Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH, UK

Received January 4, 1982. Accepted February 3, 1982.

We describe the use of a synthetic primer to select a cDNA recombinant clone containing H5 coding sequences. The strategy used was as follows: 1. Prepare oligo(dT) cellulose-bound mRNA from chicken reticulocytes and select 11S–18S material from sucrose gradients. 2. Use this RNA fraction both to prepare a cDHA library and as a template for H5-specific cDNA synthesis using a synthetic primer. 3. Screen out most globin cDNA recombinants with oligo(dT)-primed globin cDNA. 4. Search for H5 recombinants using H5 specific cDNA and verify the identity by DMA sequencing. Our screening suggests an H5 mRNA abundance of about two parts per thousand in chicken reticulocyte poly(A)-contain1ng RNA. The isolation of an H5 cDNA recombinant clone is an initial step in the study of H5 genes and their relationship to H1 and core histone genes.


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