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Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA
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Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA

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Evidence that populations of Dictyostelium single-copy mRNA transcripts carry common repeat sequences

K L Kindle, R A Firtel

    Nucleic Acids Research
    |June 11, 1979
    PubMed

    Insights

    Two plasmids, M4 and KH10, containing Dictyostelium DNA, hybridize abundant messenger RNA (mRNA). Most hybridization occurs with short, repeated DNA sequences within the plasmids, which are transcribed into various mRNAs.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Recombinant plasmids M4 and KH10 contain Dictyostelium DNA.
    • These plasmids are complementary to abundant mRNA populations in Dictyostelium.

    Purpose of the Study:

    • To analyze the sequence organization of Dictyostelium DNA in plasmids M4 and KH10.
    • To understand the nature of mRNA hybridization to these recombinant plasmids.

    Main Methods:

    • Construction of recombinant plasmids using poly(dA)-poly(dT) tailing.
    • Hybridization analysis of plasmids with pulse-labeled poly(A)+ RNA from Dictyostelium.
    • DNA sequence analysis to determine the organization of inserted Dictyostelium DNA.

    Main Results:

    • Both M4 and KH10 plasmids hybridize a heterogeneous class of mRNAs.
    • KH10 hybridizes 5-10% of vegetative cell poly(A)+ RNA.
    • Dictyostelium DNA in plasmids consists of mostly single-copy sequences interspersed with short repeated DNA sequences.
    • Hybridization is primarily to these short repeated DNA sequences.
    • M4 DNA contains a sequence complementary to a discrete mRNA.

    Conclusions:

    • The abundant mRNA populations complementary to M4 and KH10 hybridize mainly to repeated DNA sequences within the plasmids.
    • These repeated DNA sequences are transcribed into diverse single-copy mRNAs in the Dictyostelium genome.

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