Difference between revisions of "Part:BBa K5439003"

Revision as of 23:05, 1 October 2024

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.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 1947
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 2178
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 2833

@@ Line 11: / Line 11: @@
-__TOC__
-=Usage and Biology=
-<div style="text-align:justify;">
 {| class="wikitable" style="margin:auto; text-align:center; length: 80%"
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 |}
+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 <b>Figure 3</b>. The total vector concentration was 100 nanograms, whereas the reaction volume was 20 µL. Next, <b>Table 2</b> displays the protocol followed for the reaction.
+{| class="wikitable" style="margin:auto; text-align:center; length: 80%"
+|+ Table 2. DNA ligation conditions.
+|-
+!Reactive !! Quantity
+|-
+| style="text-align:center;" style="width: 80%;" | T4 DNA Ligase Buffer (10X) || 2 µL
+|-
+| style="text-align:center;" style="width: 80%;" | Vector DNA || 100 ng
+|--
+| style="text-align:center;" style="width: 80%;" | Insert DNA || 773.5 ng
+|-
+| style="text-align:center;" style="width: 80%;" | Nuclease-free water || To 20 µL
+|-
+| style="text-align:center;" style="width: 80%;" | T4 DNA Ligase || 1.5 µL
+|}
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 <span class='h3bb'>Sequence and Features</span>