A full-length cDNA clone, BHL4-1, encoding factor B was isolated from a human liver cDNA library and sequenced in its entirety. It consists of 2388 bp which include a 5'-untranslated region of 40 bp, a single open reading frame, 2292 bp in length, and a 3'-untranslated region of 56 bp followed by a poly-A tail. The deduced amino acid sequence comprises 25 residues of a putative leader peptide and 739 residues of the mature polypeptide chain of the F allele of factor B. We constructed an S allele-like Q7R mutant of BHL4-1 by site-directed mutagenesis. Both the wild-type and mutant factor B cDNA were expressed transiently in a eukaryotic system. The specific hemolytic activities of the two recombinant factor B alleles and of native B were not significantly different from each other.