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Cloning and expression of human ciliary neurotrophic factor
A Negro1, E Tolosano, S D Skaper
1Advanced Technology Division, Abano Terme, Italy.
European Journal of Biochemistry
|October 1, 1991
Summary
Researchers cloned the human Ciliary Neurotrophic Factor (CNTF) gene, finding it
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Ciliary neurotrophic factor (CNT) is a known survival factor for specific neuronal cell types in vitro.
- Its precise role in the nervous system physiology and pathology requires further investigation.
Purpose of the Study:
- To clone the entire genomic sequence of human Ciliary Neurotrophic Factor (CNTF).
- To determine the primary structure of human CNTF.
- To express biologically active human CNTF.
Main Methods:
- Cloning of the entire human CNTF gene.
- Expression of CNTF in Chinese hamster ovary (CHO) cells.
- Sequence analysis and comparison with other species.
Main Results:
- The complete genomic sequence and primary structure of human CNTF were determined.
- Biologically active human CNTF was successfully expressed in CHO cells.
- Human CNTF shows no significant sequence similarity to other known proteins but shares 84% similarity with rat and rabbit CNTF.
- Evidence suggests human CNTF is a cytosolic protein, lacking a signal sequence and glycosylation sites, and is not secreted.
Conclusions:
- The cloning and characterization of human CNTF provide a foundation for understanding its function.
- The cytosolic nature of CNTF suggests intracellular mechanisms of action.
- Further research can explore CNTF's role in nervous system health and disease.