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Cloning and expression of a cDNA encoding ovine interleukin 7
G J Barcham1, A E Andrews, A D Nash
1Centre for Animal Biotechnology, School of Veterinary Science, University of Melbourne, Parkville, Australia.
Gene
|March 10, 1995
Summary
Researchers successfully amplified ovine interleukin-7 (IL-7) cDNA, confirming its biological activity in stimulating ovine thymocyte proliferation. This finding advances understanding of immune responses in sheep.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Interleukin-7 (IL-7) is a crucial cytokine for lymphocyte development.
- Understanding ovine IL-7 is important for comparative immunology and veterinary research.
Purpose of the Study:
- To clone and characterize ovine interleukin-7 (IL-7) cDNA.
- To confirm the biological activity of the expressed ovine IL-7 protein.
Main Methods:
- Polymerase chain reaction (PCR) amplification of ovine IL-7 cDNA from activated ovine lymph node cells.
- Nucleotide and amino acid sequence analysis and comparison with human and murine IL-7.
- Expression of ovine IL-7 in COS and CHO-K1 cells and assessment of its biological activity on ovine thymocytes.
Main Results:
- Ovine IL-7 cDNA was successfully amplified and sequenced, showing homology to human and murine IL-7.
- The cDNA encodes a 176-amino acid polypeptide, processed to a 151-amino acid protein with IL-7 biological activity.
- Recombinant ovine IL-7 synergized with suboptimal concanavalin A and recombinant human IL-2 to induce ovine thymocyte proliferation.
Conclusions:
- Ovine IL-7 cDNA has been cloned and functionally characterized.
- The expressed ovine IL-7 protein demonstrates biological activity, promoting ovine thymocyte proliferation.
- Ovine IL-7 plays a role in T-cell development and can synergize with other cytokines like IL-2.