The use of single strand conformation polymorfism analysis for the study of the mitochondrial ATP-ase 6 gene.

PCR-SSCP analysis has already proven to be a simple and sensitive method for the detection of mutations in DNA. The method is based on the observation of alterations in the mobility of defined DNA fragments due to sequence variation(s).
At present we are using the PCR-SSCP technique to obtain a more detailed analysis of the mtDNA of patients suspected of mitochondrial encephalomyopathy.
A sequence examination of several patients by different research groups already showed a point mutation at nucleotide 8993 in the mitochondrial ATP-ase 6 gene.
Three partially overlapping PCR amplification fragments resulting in products of respectively 268 bp, 331 bp and 278 bp, are synthesized for a reliable screening of the whole ATP-ase 6 gene.
To avoid misinterpretation of natural polymorphisms in the ATP-ase 6 gene, observed by PCR-SSCP mobility, we are also screening normal and unrelated control persons. Human mtDNA is highly polymorphic and already 8 functionally tolerated sequenced variants (giving rise to aa variants) were reported (in the ATP-ase 6 gene). In the long run the collected normal polymorphic PCR-SSCP variant data will provide us with a solid reference base for the detection of disease-related mutations in the ATP-ase 6 gene of patients with mitochondrial encephalomyopathy.