Part:BBa_K5439003

FRET-based system for the detection of rifampicin

FRET-based sensor system for the detection of rifampicin that consists of rifampicin monooxygenase (K4447003), an enzyme that catalyzes the hydroxylation of rifampicin, flanked by two fluorescent proteins: ECFP (BBa_K1159302) as energy donor and mVenus (BBa_K1907000) as an energy acceptor.

With the DNA fragments purified from an agarose gel, we performed ligation at a molar ratio of 1:5 for vector and insert, as shown in Figure 3. The total vector concentration was 100 nanograms, whereas the reaction volume was 20 µL. Next, Table 2 displays the protocol followed for the reaction.

Table 2. DNA ligation conditions.

Reactive	Quantity
T4 DNA Ligase Buffer (10X)	2 µL
Vector DNA	100 ng
Insert DNA	773.5 ng
Nuclease-free water	To 20 µL
T4 DNA Ligase	1.5 µL