The Effect Of Mitochondrial DNA Mutation On The Penetrance And Visual Acuity Of Leber Hereditary Optic Neuropathy

Objective:To investigate the effect of mtDNA mutation on the penetance and visual acuity, 7 pedigrees of LHON were studied with pedigrees analysis, DNA sequencing and denaturing high-performance liquid chromatography (dHPLC) analysis.Material and methods:7 LHON pedigrees from (101 maternal members) Henan Province were given a standard ophthalmological exam by experienced ophthalmologist. Venous blood samples were obtained from the prohands, 96 maternal members and 20 family members, and total genomic DNA was extracted from their buffy coats by standard phenol-chloroform techniques. Individuals' DNA fragment spanning the position G3460A, G11778A, T14484C were PCR(polymerase chain reaction)-amplified and DNA sequenced. Construct four kinds of recombinant plasmid with normal or mutant DNA fragment spanning the position G3460A, G11778A . In order to get enough recombinant-plasmids, we cultured four kinds of bacterial clone with one of recombinant-plasmids respectively. Mixed two kinds of recombinant- plasmids with the same position at different proportion, then were amplified by PCR and incised by restriction enzyme. The dHPLC analysis was performed on a WAVE DNA Fragment...