Figure 1.

ATP7A transcripts in fibroblasts from a heterozygous female with the mutation c.1554-?_2172 +?del (Ex6_9del). Clone 43 which is characterized by increased copper accumulation and retention (63.4 ng ⁶⁴Cu/mg protein/20 hours and 61.1% retention) expresses only the mutant transcripts (1365 bp, missing exon 6-9). In contrast clone 29, characterized by normal copper uptake and retention (11.2ng ⁶⁴Cu/mg protein/20 hours and 11.3% retention) expresses only wild-type transcripts (1880 bp). In uncloned cell populations with normal copper uptake and retention (23.1 ng ⁶⁴Cu/mg protein/20 hours and 11.3% retention) from the same female, only normal wild-type transcripts were detected. PCR amplification of a cDNA fragment from exon 4 to exon 10 was performed by nested PCR with the primer-pair (5'-caaaagcagcccaagtacctc-3'/5'-ggtggttgccagcacaatcagt-3') followed by the primer-pair (5'-cagaagggtcacagcaaagg-3/'5'-ggtggttgccagcacaatcagt-3). I: index (D98-34147H), affected male with the mutation Ex6_9del in ATP7A. C: a control sample (D98-35541H). Pool: uncloned pool from the heterozygous female (D98-35540H). Cl43: Clone 43. Cl29: Clone 29. The PCR products were separated on a 2% agarose gel and visualized with ethidium bromide.