Plasmid and stable cell linereconstruction of RET point mutation in medullary thyroid carcinoma pedigree / 中国耳鼻咽喉头颈外科

ABSTRACT

OBJECTIVEThe plasmid construction and validation of RET point mutation in vitro.METHODS The RET gene exon in the mainland familial medullary thyroid carcinoma family was analized with muon capture two generation sequencing of. In vitro, the RET relative point mutationswere reconstructed in NIH3T3 cells and the expression levels were studied.RESULTSThe corresponding lentiviral plasmids of RET point mutations were successfully obtained after point mutating the wild RET plasmids, it was verified that the target genes were expressed in NIH3T3 cells stably by Western Blot. CONCLUSIONSStable cell lines carrying RET point mutations were reconstructedsuccessfully, which provide a good platform for studying various point mutations.