Difference between revisions of "Part:BBa K1316008"

Revision as of 14:10, 15 October 2014

ybiJ promoter coupled to mKate2 reporter gene and Rhamnose promoter coupled to N-genes

Promoter of the ybiJ gene, which is activated by the presence of several nitrogen-based compounds such as 2,4,6-TNT, 2,4-DNT and 1,3-DNB. mKate2 is a far-red fluorescent protein used with reporting purposes. mKate2 codon usage is optimised for high expression in mammalian cells (humanised). It is, nevertheless, suitable for propagation in E. coli. nfsA, nfsB and nemA (refered here as N-genes) are the genes for NfsA, NfsB and NEM reductases, which presumably play a role in 2,4-DNT and 2,4,6-TNT metabolism. These genes are regulated by the inducible Rhamnose promoter This construct is, then, designed to be able to detect and quantify the presence of compunds such as 2,4,6-TNT, 2,4-DNT and 1,3-DNB. The presence of the N-genes aims to enhance this response.

Characterisation

Adding these genes did unfortunately show no clear improvement on the response of the ybiJ promoter in front of 2,4-DNT.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 381
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 886
Illegal BsaI.rc site found at 1075

@@ Line 6: / Line 6: @@
 nfsA, nfsB and nemA (refered here as N-genes) are the genes for NfsA, NfsB and NEM reductases, which presumably play a role in 2,4-DNT and 2,4,6-TNT metabolism. These genes are regulated by the inducible Rhamnose promoter
 This construct is, then, designed to be able to detect and quantify the presence of compunds such as 2,4,6-TNT, 2,4-DNT and 1,3-DNB. The presence of the N-genes aims to enhance this response.
+<h3>Characterisation</h3>
 Adding these genes did unfortunately show no clear improvement on the response of the ybiJ promoter in front of 2,4-DNT.