[in Japanese]

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The complexes AP_ν (A: bovine serum albumin (BSA), P: palmitic acid, ν: 0.63-8.1) were exposed to high pH's for various periods (15min.-7h) at 20°C. The results obtained by the quantitative analysis of disc gel electrophoresis are as follows: 1. When the complexes were exposed to pH 11.3 for 1h, the BSA containing 6-8mols of palmitic acid per mol of albumin (AP₆-AP₈) did not cause the “first reaction” and was not denatured at all. 2. With further increase in exposure pH and/or exposure time, the “first reaction” progressed over various compositions of the complex. Namely, there was no resistant form to alkaline denaturation under these conditions of exposure; nevertheless, AP₆-AP₈ revealed the higher stability than any other complex against the denaturation.

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We have studied two patients (case 1 and case 2) with unstable lactate dehydrogenase (LDH) H(B) subunit. We previously detected a single base substitution of G by A at Arg codon CGC in exon 4 of case 1 and of A by C at Ser codon AGT in exon 3 of case 2. These mutations result in an amino acid replacement of a conserved Arg by His at the residue “173” and Ser by Arg at the residue “131”, respectively We designed mismatch primers for the detection of the point mutations in exon 4 and exon 3, respectively. These mismatched PCR methods could be easily and clearly detect these missense mutations. Genomic analysis of case 2 family by the mismatched PCR showed that the missense mutation was consistent with their biochemical phenotypes. The mismatched PCR does not require radioisotopes or a cumbersome hybridization procedure, and can be easily performed any molecular diagnosis. A simple and convenient method maily using electrophoresis has been established.

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