Molecular analysis.

(A) Direct sequencing of the amplified CPO genomic DNA fragment from the patient. The sequence around the mutation site is shown. A single base substitution, G-to-A, at the last nucleotide of the splicing donor site of exon 6 is shown as double peaks and is indicated by an arrow. (B) Pedigree of the family. Circles denote females; squares denote males; half-shaded symbols denote HCP heterozygous. N indicates not tested. An arrow denotes the proband MspI restriction site analysis of the amplified genomic DNA fragment. PCR-amplified fragments (232 bp) of the mother (lane 1) and a healthy control not related to the family (C) were completely digested into 182-bp and 50-bp fragments, whereas those of the patient (lane 2) and the father (lane 3) were only partially digested. Lane M, DNA molecular weight standard pHY size marker (Toyobo, Tokyo, Japan). Numbers on the left and right indicate the sizes of DNA size markers and fragments, respectively, observed in this analysis. (C) Sequence around the region encoding the amino acid Lys 404 showed a normal pattern.