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Molecular cloning of human interferon cDNA
Abstract:
A hybrid plasmid, TpIF319, has been shown to contain the sequence for human fibroblast interferon mRNA [Taniguchi, T., Sakai, M., Fujii-Kuriyama, Y., Muramatsu, M., Kobayashi, S. & Sudo, T. (1979) Proc. Jpn. Acad. 55, Ser. B, 464-469]. This conclusion was confirmed by a hybridization-translation assay, using rabbit globin mRNA and its cDNA-containing plasmid as a control. Plasmid TpIF319 was used as probe and another recombinant plasmid, TpIF319-13, whose cDNA insert consists of about 800 base pairs, was isolated. Nucleotide sequence analysis of the cDNA revealed that the DNA in fact codes for human fibroblast interferon.
Insights
Researchers confirmed a hybrid plasmid, TpIF319, contains the human fibroblast interferon mRNA sequence. Further analysis of a derived plasmid, TpIF319-13, verified the DNA sequence codes for this important human interferon.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- The identification and characterization of specific gene sequences are crucial for understanding cellular functions and developing therapeutic agents.
- Human fibroblast interferon plays a significant role in the immune response and has therapeutic potential.
Purpose of the Study:
- To confirm that the hybrid plasmid TpIF319 contains the sequence for human fibroblast interferon mRNA.
- To isolate and characterize a recombinant plasmid with a substantial cDNA insert for detailed sequence analysis.
Main Methods:
- Hybridization-translation assays were employed to verify the presence of interferon mRNA sequences.
- Recombinant plasmid isolation and purification were performed.
- Nucleotide sequence analysis of the isolated cDNA insert was conducted.
Main Results:
- The hybrid plasmid TpIF319 was confirmed to contain the human fibroblast interferon mRNA sequence.
- A recombinant plasmid, TpIF319-13, with an approximately 800 base pair cDNA insert was successfully isolated.
- Sequence analysis definitively showed that the cDNA insert codes for human fibroblast interferon.
Conclusions:
- The nucleotide sequence analysis provides conclusive evidence that the isolated DNA fragment codes for human fibroblast interferon.
- This study validates the use of plasmid TpIF319 as a source for human fibroblast interferon genetic information.
- The findings contribute to the understanding of interferon gene structure and facilitate further research in interferon-based therapeutics.