Team:Wisconsin-Madison/parts 1

Taq Ligase

Taq Ligase is a thermostable DNA ligase, which joins adjacent 5' and 3' ends of DNA, removing nicks in double stranded DNA, active from 45-65 degrees Celsius(Barany, F. (1991). Proc. Natl. Acad. Sci. USA. 88, 189-193). The Taq ligase gene is within a kanamycin resistant PET-ALM1 plasmid, downstream of the T7 promoter, upstream of the T7 terminator. Thus, it can be used with an inducible T7 expression system to express the enzyme. The protein has an N-Terminus hexa-histidine tag, joined to the wild type structure by a thrombin cleavage motif. Note: our observation was that expression of Taq Ligase was semi toxic to cells, causing significantly slower growth of cultures containing this plasmid.