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Sequence and expression of rat ICAM-1.
1Molecular Biology Research Laboratory, Tokyo R and D Center, Daiichi Pharmaceutical Co., Japan.
Biochimica Et Biophysica Acta
|May 7, 1992
Summary
Researchers isolated rat intercellular adhesion molecule-1 (RICAM-1) cDNA, finding significant homology with mouse and human ICAM-1. Transfected CHO cells expressed high levels of RICAM-1 on their surfaces.
Area of Science:
- Molecular Biology
- Immunology
- Cell Biology
Background:
- Intercellular Adhesion Molecule-1 (ICAM-1) plays a crucial role in immune responses and cell adhesion.
- Understanding the molecular characteristics and expression of ICAM-1 in different species is vital for immunological research.
Purpose of the Study:
- To isolate and characterize the cDNA for rat intercellular adhesion molecule-1 (RICAM-1).
- To investigate the expression of RICAM-1 in a heterologous cell system.
Main Methods:
- Construction of a cDNA library from IL-1 beta-stimulated rat Ax cells.
- Hybridization screening using mouse ICAM-1 cDNA.
- Nucleic acid sequence homology analysis.
- Construction of a transfection vector (pSV-RICAM1-neo) with the SV40 promoter.
- Flow cytometric analysis of transfected Chinese hamster ovary (CHO) cells.
Main Results:
- Isolation of cDNA clones encoding RICAM-1.
- RICAM-1 exhibits 79.1% nucleic acid homology with mouse ICAM-1 and 55.6% with human ICAM-1.
- Transfection of CHO-K1 cells with pSV-RICAM1-neo resulted in high-level surface expression of RICAM-1.
Conclusions:
- The study successfully isolated and characterized RICAM-1 cDNA.
- RICAM-1 shares significant sequence homology with its murine and human counterparts.
- Engineered CHO cells can effectively express RICAM-1, providing a model for further functional studies.