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A signal sequence trap based on a constitutively active cytokine receptor
1Department of Hematopoietic Factors, The Institute of Medical Science, University of Tokyo, Japan.
Nature Biotechnology
|May 20, 1999
Summary
Researchers developed a novel method to identify signal sequences in cDNA fragments. This technique enables the discovery of genes encoding secreted and cell-surface proteins, including type II membrane proteins, using interleukin-3 (IL-3)-independent cell growth.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Signal sequences direct proteins to the cell membrane.
- Existing methods for signal sequence detection have limitations.
Purpose of the Study:
- To develop a new method for detecting signal sequences in cDNA fragments.
- To identify novel genes encoding secreted and cell-surface proteins.
Main Methods:
- A functional assay using a cytokine receptor and interleukin-3 (IL-3)-dependent Ba/F3 cells.
- Retrovirus-mediated gene transfer and selection for IL-3-independent growth.
- Analysis of isolated cDNA clones for encoded proteins.
Main Results:
- The method successfully identified known and novel cDNA clones.
- All known clones encoded secreted and cell-surface proteins.
- Type II membrane proteins, previously undetected, were isolated.
Conclusions:
- The developed method is effective for identifying signal sequences.
- This approach expands the discovery of secreted and cell-surface proteins, including type II membrane proteins.