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cDNA of a novel mRNA expressed predominantly in mouse kidney
T Kawamura1, N Kuroda, Y Kimura
1Department of Molecular Biology, Nagoya City University School of Medicine, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan.
Abstract:
We examined embryonic carcinoma (EC) cells for a potential prototype molecule of C3, the third component of complement. PCR primers, corresponding to the base sequence derived from the C3 cDNA of several species, were used for PCR amplification of the EC cell cDNA. All the PCR products obtained had the same sequence and showed no sequence homology to C3. Subsequently, cDNA clones were isolated from a mouse liver cDNA library using the PCR product as a probe. Unexpectedly, neither the base sequence of the cDNA clones nor the amino acid sequence deduced from the cDNA showed homology to C3, although partial homology was observed to a number of sequences from EST databases. We designated this new clone NCU-G1. Northern hybridization experiments revealed that NCU-G1 is expressed constitutively not only in the mouse fetus but also in various mouse tissues, and is most abundant in the kidney cortex.
Insights
Researchers investigated embryonic carcinoma cells for a C3 complement prototype molecule. They discovered a novel gene, NCU-G1, unrelated to C3, expressed widely in mouse tissues, particularly the kidney.
Area of Science:
- Molecular Biology
- Immunology
- Genetics
Background:
- The complement system, particularly C3, plays a crucial role in innate and adaptive immunity.
- Embryonic carcinoma (EC) cells are pluripotent cells with potential for differentiation studies.
Purpose of the Study:
- To identify a potential prototype molecule of C3 within embryonic carcinoma (EC) cells.
- To characterize a novel gene discovered during the investigation.
Main Methods:
- Polymerase Chain Reaction (PCR) amplification using primers derived from C3 cDNA.
- cDNA library screening using PCR products as probes.
- DNA sequencing and homology analysis.
- Northern hybridization for gene expression analysis.
Main Results:
- PCR amplification of EC cell cDNA did not yield sequences homologous to C3.
- A novel cDNA clone, designated NCU-G1, was isolated and characterized.
- NCU-G1 showed no homology to C3 but exhibited partial homology to sequences in EST databases.
- Northern hybridization revealed constitutive expression of NCU-G1 in fetal and adult mouse tissues, with highest abundance in the kidney cortex.
Conclusions:
- The investigated EC cells do not appear to express a C3 prototype molecule.
- A novel gene, NCU-G1, has been identified and its expression pattern characterized.
- NCU-G1 represents a new molecular entity with potential roles in kidney function or development.