Analysis of PKD2 gene variant and protein localization in a pedigree affected with polycystic kidney disease.
10.3760/cma.j.cn511374-20191204-00619
- Author:
Jianping CHENG
1
;
Ping LI
;
Yujun LI
;
Yong'an ZHOU
;
Ruirui REN
;
Yaxin HAN
;
Xingxing LI
;
Zhe LI
;
Yuan BAI
Author Information
1. Shanxi Medical University Graduate School, Taiyuan, Shanxi 030001, China. zya655903@163.com.
- Publication Type:Journal Article
- MeSH:
DNA Mutational Analysis;
Female;
Frameshift Mutation;
HEK293 Cells;
HeLa Cells;
Humans;
Male;
Pedigree;
Polycystic Kidney, Autosomal Dominant/physiopathology*;
Protein Kinases/genetics*;
Protein Transport/genetics*;
Whole Exome Sequencing
- From:
Chinese Journal of Medical Genetics
2021;38(1):47-51
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To detect the mutation site in a pedigree affected with autosomal dominant polycystic kidney disease (ADPKD) and verify its impact on the protein function.
METHODS:Peripheral blood samples were collected from the proband and his pedigree members for the extraction of genomic DNA. Mutational analysis was performed on the proband through whole-exome sequencing. Suspected variant was verified by Sanger sequencing. A series of molecular methods including PCR amplification, restriction enzyme digestion, ligation and transformation were also used to construct wild-type and mutant eukaryotic expression vectors of the PKD2 gene, which were transfected into HEK293T and HeLa cells for the observation of protein expression and cell localization.
RESULTS:The proband was found to harbor a c.2051dupA (p. Tyr684Ter) frame shift mutation of the PKD2 gene, which caused repeat of the 2051st nucleotide of its cDNA sequence and a truncated protein. Immunofluorescence experiment showed that the localization of the mutant protein within the cell was altered compared with the wild-type, which may be due to deletion of the C-terminus of the PKD2 gene.
CONCLUSION:The c.2051dupA (p. Tyr684Ter) mutation of the PKD2 gene probably underlay the pathogenesis of ADPKD in this pedigree.
