Figure 5

Novel in-frame deletion (L3287del) identified in family PK002. A. MRF-SSCP analysis of SI 7 fragment of the patient. The restriction map shows multiple fragment sizes obtained from digestions with two sets of restriction endonucleases (upper). The dotted square indicates the region with mobility shifts in the digested SI 7 fragment from the patient of family PK002. The SSCP patterns generated from digestions with two sets of enzymes (lower) and mobility shifts of single-stranded DNA (pointed) are shown. B. Direct sequencing analysis of PKD1 -cDNA from the patient. The sequencing profile shows a deletion of three nucleotides (CTC; c.10070_10072del), resulting in an in-frame mutation. The CTC deletion causes missing of leucine at position 3287 in polycystin-1 (L3287del). C. Multiple sequence alignment of polycystin-1 from human, dog, rat, mouse, pufferfish and worm by using ClustalW program [33]. Amino acid residues around L3287 are shown. L3287 is highly conserved among distant species in which only in C. elegans that the residue is replaced by isoleucine, which is in the same group of amino acid. D. Direct detection for L3287del in members of the family PK002 by SSCP analysis. The L3287del could be detected by the presence of heteroduplex DNAs on the SSCP analysis. In the family PK002, five members carried the mutation while nine did not. M is standard DNA markers and ds is double-stranded DNA of a normal control.