The Expression And Function Of Mutants Of SEDT-PA'S Pathogenic Gene WISP3 In Chondrocytes

Part One The Construction of Mutants of SEDT-PA's Pathogenic GeneObjectives: Wnt-inducible secreted protein 3 (WISP3) was spondyloepiphyseal dysplasia tarda with progressive arthropathy (SEDT-PA)'s pathogenic gene.To construct two types of WISP3 gene's mutants (1000T-C,840delT) found in SEDT-PA patients.Methods: Full-length cDNAs of wild type WISP3 gene(WT-WISP3) was amplified from human chondrocytes by RT-PCR, and site-directed mutagenesis was used to obtain full-length cDNAs of the mutated WISP3 genes (MUT^1000T/C and MUT^840delT) . The products were cloned into pEGM-T-easy vector and their sequences were analysed, then subcloned into eukaryotic expression vector pcDNA3.1 (+) and pEGFP-C2, the inserted fragments were analysed and reconfirmed by restriction endonuclease analysis and sequencing.Results: By restriction endonuclease analysis and sequencing, the sequences of WT-WISP3, MUT^1000T/C and MUT^840delT were consistent with that in literature and mutated in SEDT-PA, and the open reading frames were matched with the vector sequence.Conclusion: WISP3 gene's mutants of SEDT-PA were successfully...