A novel PCR-based technique using expressed sequence tags and gene homology for murine genetic mapping: localization of the complement genes.
The complement system is a cascade of serum proteins and receptors which forms a vital arm of innate immunity and enhances the adaptive immune response. This work establishes the chromosomal localization of four key genes of the murine complement system. Mapping was performed using a novel and rapid PCR restriction length polymorphism method which was developed to exploit the murine expressed sequence tag (EST) database. This technique circumvents the laborious cDNA or genomic cloning steps of other mapping methods by relying on EST data and the prediction of exon-intron boundaries. This method can be easily applied to the genes of other systems, ranging from the interests of the individual researcher to large-scale gene localization projects. Here the complement system, probably one of the most well-characterized areas of immunology, was used as a model system. It was shown that the C3a receptor C1r and C1s genes form an unexpected complement gene cluster towards the telomeric end of chromosome 6. The second mannose binding lectin-associated serine protease gene was mapped to the telomeric end of chromosome 4, which is distinct from other complement-activating serine proteases. These results provide new insights into the evolution of this group of proteins.
Lawson
P R
PR
MRC Immunochemistry Unit, Department of Biochemistry, South Parks Road, Oxford University, Oxford OX1 3QU, UK.
Reid
K B
KB
D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D002877:Chromosomes, Human / Q000235:genetics; D015923:Complement C1r / Q000235:genetics; D003173:Complement C1s / Q000235:genetics; D003165:Complement System Proteins / Q000235:genetics*; D019143:Evolution, Molecular; D020224:Expressed Sequence Tags; D006801:Humans; D050606:Mannose-Binding Protein-Associated Serine Proteases; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D009115:Muridae / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011951:Receptors, Complement / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics; D013045:Species Specificity; D016615:Telomere / Q000235:genetics
D008565:Membrane Proteins; D011951:Receptors, Complement; C085567:complement C3a receptor; D003165:Complement System Proteins; C105296:MASP2 protein, human; D050606:Mannose-Binding Protein-Associated Serine Proteases; D012697:Serine Endopeptidases; D015923:Complement C1r; D003173:Complement C1s