Streamlined Approach to Functional Analysis of Promoter-Region Polymorphisms

S.L. Coleman, B. Hoogendoorn, C. Guy, S.K. Smith, M.C. O'Donovan, P.R. Buckland
2002 BioTechniques  
We have developed a rapid method for identifying functional promoter-region polymorphisms. Using a modified pGL3 luciferase expression T-vector, we can amplify by PCR, clone, identify allelic pairs of a polymorphic gene promoter region, and prepare plasmids for cell culture 10 promoters (20 allele pairs) per week per researcher. By utilizing 96-well plate technology and an internal control plasmid expressing secreted alkaline phosphatase, each of these allele pairs can be tested for relative
more » ... moter activity in each of three cell lines (HEK293t, TE671, and JEG3) with similar resources.
doi:10.2144/02332ht01 pmid:12188194 fatcat:nu2le4l5uncezbghgaa6ubqqby