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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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Related Experiment Video

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Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
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The primary sequence of rabbit alpha-globin mRNA

H C Heindell, A Liu, G V Paddock

    Cell
    |September 1, 1978
    PubMed
    Summary

    Researchers have fully sequenced the rabbit alpha-globin messenger RNA (mRNA), confirming its accuracy through complementary RNA (cRNA) sequencing. This detailed sequence provides insights into gene expression and protein synthesis.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • The rabbit alpha-globin gene is crucial for hemoglobin production.
    • Accurate sequencing of messenger RNA (mRNA) is essential for understanding gene expression and protein synthesis.

    Purpose of the Study:

    • To determine the complete nucleotide sequence of rabbit alpha-globin messenger RNA (mRNA).
    • To validate the accuracy of the sequenced mRNA through comparison with existing data and complementary RNA (cRNA) sequencing.
    • To analyze the general features of the mRNA sequence, including base composition and codon preferences.

    Main Methods:

    • DNA sequencing using the Maxam and Gilbert method.
    • Analysis of the chimeric plasmid pHb 72 containing the rabbit alpha-globin DNA insertion.

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  • Comparison of obtained mRNA sequence with previously published data for 5' and 3' ends.
  • Utilizing a new computer program to analyze base-pairing arrangements.

    • Main Results:

    • The complete primary sequence of rabbit alpha-globin mRNA was determined, spanning the 5' untranslated region through the first 361 translated nucleotides.
    • The determined sequence showed complete agreement with previously reported overlapping sequences.
    • The results reaffirmed the high fidelity of in vitro synthesized gene copies, bacterial cloning, and maintenance.

    Conclusions:

    • The complete rabbit alpha-globin mRNA sequence has been established with high fidelity.
    • The findings support the reliability of synthesizing, cloning, and maintaining gene copies in vitro.
    • Analysis of the sequence provides valuable data for understanding gene expression, codon usage, and mRNA structure.