Fig. 7
From: Engineered bidirectional promoters enable rapid multi-gene co-expression optimization
Bidirectional transcription terminators (BDTs) required for the assembly of bidirectional multi-gene co-expression relieve expression loss associated with transcriptional collision. A reporter construct for testing bidirectional transcription termination was assembled by cloning the genes coding for eGfp and dTom in convergent orientation (small inlet). Two AOX1 promoters were used to drive equal expression of the reporter genes. Monodirectional terminators (MDTs) were combined into bidirectional fusion terminators and two putative natural BDTs (nBDTs) were tested. A negative control lacking termination sequences and bearing solely a NotI restriction site was included. Additional control constructs contain only a single AOX1 promoter, a single FP, and the AOX1* terminator. AOX1TT* denotes the AOX1 terminator sequence used by Vogl et al.30. Some BDTs acted also as autonomously replicating sequences (Supplementary Fig. 7). Mean values and standard deviations of fluorescence measurements after pre-growth on glucose followed by methanol induction of biological quadruplicates are shown
