Transcription-coupled repair pathway in UVC-induced SupF gene mutation in Tet-on 293 cell line⋆

ObjectiveTo explore the role of transcription-coupled repair (TCR) pathway in the UVC-induced SupF gene mutation in Tet-on 293 cell line, we designed and constructed a Tet-responsive plasmid DNA pTCR-C1, and utilized this pTCR-C1 plasmid to obtain the mutation frequency of SupF reporter gene induced by UVC in Tet-on 293 cell line.MethodsSupF gene was cloned into a Tet-responsive plamid pBI-L, which include a bidirectional Tet-responsive promoter, and was named pTCR-C1. The pTCR-C1 plasmid was transfected into Tet-on 293 cell line, and the mutation frequency of SupF reporter gene was detected in the presence and absence of DOX.ResultsThe pTCR-C1 plasmid was identified with the methods of restriction digestion and DNA sequencing. The mutation frequency of SupF reporter gene in the presence of DOX was higher than in the absence of DOX.ConclusionThe TCR pathway takes part in the UVC-induced SupF gene mutation in Tet-on 293 cell line.