Related Experiment Videos
Identification of a protein encoded by a mouse simple repeated sequence
M Di Carlo1, D P Romancino, G Ghersi
1Institute of Developmental Biology, C.N.R., Palermo, Italy.
Journal of Submicroscopic Cytology and Pathology
|April 1, 1994
Summary
Researchers explored if mouse simple sequence DNA could encode proteins. They found a 28 KDa protein antigen in mouse tissues, detectable in the cytoplasm using specific antibodies.
Area of Science:
- Molecular Biology
- Protein Expression
- Immunology
Background:
- Simple sequence DNA, including ORFs, may encode proteins.
- Investigating novel protein expression from repetitive DNA elements is crucial.
Purpose of the Study:
- To determine if a mouse simple sequence DNA fragment containing an ORF can encode a protein.
- To characterize the expressed protein and its tissue distribution in vivo.
Main Methods:
- Insertion of a cDNA fragment with 'CAGAGAGG' sequence into the pEX31C expression vector.
- Expression of a fusion protein (H5fp) in E. coli and generation of polyclonal antibodies in rabbits.
- Western blot analysis and immunolocalization in mouse tissues.
Main Results:
- The fusion protein H5fp was successfully expressed.
- Anti-H5fp antibodies recognized a 28 KDa antigen in various mouse tissues.
- The antigen was localized to the cytoplasm of examined cells.
Conclusions:
- A mouse simple sequence DNA fragment can encode a detectable protein antigen.
- The expressed protein is present in the cytoplasm of mouse tissues.
- Further structural analysis using methods like Chou and Fasman prediction can be performed.