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

Paul A. Krieg | Zendy; Allan J. Robins | Zendy; Michael J. Gait | Zendy; Richard C. Titmas | Zendy; J. R. E. Wells | Zendy

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

Author(s) -

Paul A. Krieg,

Allan J. Robins,

Michael J. Gait,

Richard C. Titmas,

J. R. E. Wells

Publication year - 1982

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/10.5.1495

Subject(s) - complementary dna , biology , primer (cosmetics) , cdna library , microbiology and biotechnology , recombinant dna , rna , gene , genetics , chemistry , organic chemistry

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 cDNA 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 DNA sequencing. Our screening suggests an H5 mRNA abundance of about two parts per thousand in chicken reticulocyte poly(A)-containing 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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