A Exclusive Mapping Analysis on the Gene Associated with X-linked Dominant Inherited Nephritis in One Pedigree

Objective To study the characteristics of mutations of gene COL4A5 encoding type Ⅳ collagen α 5 chain in one chinese pedigree with X-linked dominant inherited nephritis (Alport's syndrome,AS). Methods Genomic DNA was extracted from 35 members of the pedigree of Alport's syndrome. All of 51 exons of COL4A5 gene were amplified by PCR with the primers synthesized according to the published intron sequences of COL4A5. The PCR products were further analyzed by agarose gel electrophoresis and single strand conformation polymorphism (SSCP). The PCR products showing polymorphism were directly sequenced. Results PCR-SSCP analysis showed that 17 PCR products had abnormal mobility of single strand DNA. DNA sequencing analysis revealed 9 suspicious mutations. But these suspicious mutations were not be confirmed by inverse sequencing analysis. Conclusion The exon mutation of COL4A5 gene of this pedigree did not be found, and the mutations of COL4A5 gene may locate in the its introns.