Isolation of the Xenopus complement factor B complementary DNA and linkage of the gene to the frog MHC.
C factor B (Bf) is the key component of the C3 convertase of the alternative C pathway, and its gene resides in the class III region of the mammalian MHC. To elucidate the evolution of both the C system and the MHC, we isolated Bf cDNA clones from Xenopus laevis, an ectothermic vertebrate in which the MHC has been well defined at both the biochemical and functional levels. A part of the serine protease domain of the Xenopus Bf mRNA was amplified by reverse transcriptase-PCR, using degenerate primers corresponding to regions encoding the perfectly conserved amino acid sequences found in both the mouse Bf and C2 proteins. A full length Xenopus Bf cDNA clone was isolated from a Xenopus liver cDNA library. The deduced amino acid sequence of 747 residues showed the same domain structure as mammalian Bf and C2: three short consensus repeat domains, a von Willebrand domain and a serine protease domain. Xenopus Bf has 40% and 30% overall amino acid identity to mouse Bf and mouse C2, respectively. Because the amino acid identity between mouse Bf and mouse C2 is 38%, the gene duplication of Bf/C2 probably occurred before the divergence of amphibians and mammals. Southern blotting analysis of the Xenopus Bf gene showed a close linkage to the MHC, indicating that the Bf gene was linked to the class I and class II genes at the time Xenopus shared a common ancestor with mouse and man, 350 x 10(6) yr ago.
Kato
Y
Y
Department of Biochemistry, Nagoya City University Medical School, Japan.
Salter-Cid
L
L
Flajnik
M F
MF
Kasahara
M
M
Namikawa
C
C
Sasaki
M
M
Nonaka
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D011415:Complement Factor B / Q000235:genetics*; D018076:DNA, Complementary; D008040:Genetic Linkage / Q000235:genetics*; D008285:Major Histocompatibility Complex / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014982:Xenopus laevis / Q000235:genetics* / Q000276:immunology
D018076:DNA, Complementary; D011415:Complement Factor B