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Rat complement factor I: molecular cloning, sequencing and expression in tissues and isolated cells.
G Schlaf1, E Rothermel, M Oppermann
1Department of Immunology, Georg-August University, Göttingen, Germany.
Immunology
|December 3, 1999
Summary
Researchers identified rat Factor I (FI) mRNA using PCR, revealing high sequence identity to mouse and human FI. Expression analysis showed FI mRNA is primarily found in hepatocytes and endothelial cells, not immune cells.
Area of Science:
- Biochemistry
- Immunology
- Molecular Biology
Background:
- Factor I (FI) is a crucial regulatory serine protease in the complement system, inactivating complement proteins C3b and C4b.
- Human and mouse FI are heterodimers composed of a heavy and a catalytic light chain.
Purpose of the Study:
- To clone and characterize the rat Factor I (FI) cDNA.
- To investigate the expression patterns of FI mRNA in various rat and human cell types.
Main Methods:
- Screening of a rat liver cDNA library using PCR with degenerate primers.
- Nucleotide sequence analysis and comparison with human and mouse FI sequences.
- Reverse transcription PCR (RT-PCR) assays to determine FI mRNA expression in different cell types.
Main Results:
- A rat cDNA sequence of 2243 bp was identified, containing the complete coding region and 3' untranslated region of FI.
- Rat FI coding sequence showed 87% identity to mouse and 78% to human FI at the nucleotide level.
- FI mRNA was detected in rat hepatocytes, human umbilical vein endothelial cells, rat uterus, and rat small intestine, but not in various immune cells or human lymphoma cell lines.
Conclusions:
- The rat FI sequence is highly homologous to its mammalian counterparts.
- FI expression is predominantly localized to hepatocytes and endothelial cells, suggesting specific roles beyond immune cell regulation.