Design and characterization of artificial transcriptional terminators

Abstract

Design and characterization of artificial transcriptional terminators. Ten new terminators were designed based on previous research of terminator structure and termination efficiency. The terminators were built by PCR extension, ligated into a BioBrick plasmid backbone, and transformed into TOP10 cells. Characterization devices were built to test the terminators. Input and output of the terminator were measured by expression of RFP and GFP. Charaterization devices were then placed into the E. coli strain CW2553/pJAT18, which hijacks the arabinose transport system to provide controlled input to the terminator. Of the ten terminators designed and tested, BBa_B1002, BBa_B1004, BBa_B1006 and BBa_B1010 proved to be strong terminators with termination efficiencies above 90%. These terminators may be obtained from the Registry of Standarized Parts at parts.mit.edu.