| Autosomal dominant polysystic kidney disease (ADPKD) is a single gene genetic disease mainly caused by mutations of pkd1 gene located on chromosome 16pl3.3, its pathological changes include polycystic kidney and polycystic liver in patients when growing to adult. The mutation loci in this gene from different family did not coincide with each other completely. Single nucleotide polymorphism (SNP) is a new method for genetic marker analysis. It spreads widely on chromosome and may exist in determining genes of every genetic traits. There is not pdk1 gene SNP research reported until now. pkd1 gene, with complicated structure and unclear function, is about 54kb with 46 exons and its mRNA is 12906bp. In this experiment, we carried out microsatellite DNA polymorphism analysis and mutation screening from exon 39 to 44 in 3' region oipkdl gene in collected patients, including three families and 11 no relationship individuals. Study on pkdl SNP will be helpful to elucidate its pathogenesis and disease diagnosis.The polymorphism analysis of pkdl flanking microsatellite DNA in 64 Chinese indicated that there were 11 fragments in CW2 locus ranging from 112 to 132bp, 31.25% of which were 122bp, heterozygote was 0.8271, PIC was 0.7583. There were 14 fragments in D4S1563 locus ranging from 205 to 235bp, 25% of which were 221bp, heterozygote was 0.8583, PIC was 0.8226. A special 134bp fragment of CW2, which could not be found in normal individuals, was found in ADPKD families for the first time in this experiment. By microsatellite markers analysis, the disease was linkagewith flanking microsatellites oipkdl gene in three families.The single nucleotide polymorphism oipkdl gene, the candidate gene of ADPKD patients, was analyzed. According to pkdl gene sequence in GeneBank (53522bp, Acce No. L39891), 5 pairs of primers were designed with Oligo 6.0 software. Sequence analysis was carried out from exon 39 to 44 in 3 ' end of pkdl by PCR, cloning and sequencing. Single base mutation with higher frequency spread widely in the region of exon 39 to 44, including single base transition, transversion, insertion and deletion. All this higher density mutation may be the related SNP loci of ADPKD pathogenesis. By statistic analysis, G-C and T-A transversion occuned more frequently, the ratio of transversion and transition was 5:1. The G deletion and insertion was the most which mainly located on 47721, 49387 and 48829 positions, especially on 48829 locus, which showed more complicated variations with deletion, insertion and C-G transversion.149 mutations detected in this experiment could lead to 20 missense mutations and 2 frame-shift mutations, 9 of them were new mutation found for the first time and may affect the formation of protein higher structure and function realization. The analysis on amino acid sequence oipkdl was carried out in three ADPKD families and 11 no relationship individuals. The same amino acid mutations in these patients maybe the common reason for the ADPKD pathogenesis.The single nucleotide mutations on 7 loci which caused 4 restriction enzyme sites variation were found for the first time in the study. All these variation sites could be used as candidate loci for SNPs analysis oipkdl gene and diagnosis kit development for ADPKD disease.The pathological analysis was carried out on kidney of ADPKD patients. The result showed that renal glomerulus and tubule structure changed significantly, the margin of renal cortex medulla disappeared, and the smooth muscle in blood vessel of...
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