Difference between revisions of "Part:BBa K2020052"

Revision as of 12:43, 17 October 2016

DMNBS-Synthetase for use in E.coli

This is a DMNBS-synthetase to be used as a orthogonal synthetase in E.coli. This part can be used together with the cognate tRNA BBa_K2020042 to incorporate DMNBS in response to an amber stop codon.

Compared to incorporation of tyrosine with wild type Mj Y-Synthetase in response to an amber codon:

• Incorporation Efficiency: % (DMNBS incorporation value)
• Incorporation Fidelity: %(descrimination againt tyrosine)

Full Set of DMNBS synthtetases

• DMNBS-RS Clone 1
• DMNBS-RS Clone 2
• DMNBS-RS Clone 3
• DMNBS-RS Clone 4
• DMNBS-RS Clone 5
• DMNBS-RS Clone 6
• DMNBS-RS Clone 7
• DMNBS-RS Clone 8
• DMNBS-RS Clone 9
• DMNBS-RS Clone 10

Usage and Biology

incorporation of ncAA

Assembly in a synthetase plasmid for incorporation of ncAA

alt text

Most synthetases are used with low copy plasmids (e.g. pACYC). Assemble the tRNA and the synthetase into a low copy plasmid, each one with an own promoter and one terminator for both. (See picture). If your application is not for incorporation into a protein but for use with a second plasmid, make shure to use replicons from different incompatibility groups, eg. ColE1 and p15A and different selection markers. A second plasmid could be the flourescent reporter plasmid pFRY for the purpose of determining fidelity and efficiacy of synthetases for ncAA.

Sequence and Features