Difference between revisions of "Part:BBa K549003"

Revision as of 15:21, 30 September 2011

Nickel(II) dependent promotor with luxAB as reporter

The biobrick contains the RcnA promotor from E. coli fused with the luxAB reporter gene. E. coli strains own the RcnR-regulator gene which directly regulates the transcription of the RcnA nickel-cobalt efflux protein from which the promotor is amplified. In absence of Ni(II), RcnR represses the rcnA-promoter. After addition of Ni (II) (NiCl2), RcnR binds this ion and is released from the promoter. Therefore, luxAB will be expressed.

--> also use luxCDE or luciferine to detect output.

In presence of nickel the Luciferase is expressed.

The promoter is also a little bit sensitive for Co 2+.

Usage and Biology

This is how the BioBrick is working:

The promoter sequence of pnikA is missing in the part sequence below. This is the promoter sequence of pnikA directly upstream of the reporter gene. The promoter sequence also contains the first ca. 50 nt of nikA, i.e. its a translational fusion. ACGGATTGTATGAGACATGGCAACACCTGGTTAACAAGAATATGAAAAATCATAGCACTATTAATCTACTGGGGGGTAGT ATCAGGTACTGGGGGGGAGTAGAATCAGATTGCCGAATTAATACTAAGAATTATTATCATGACCGAATTTACAACTCTTC TTCAGCAAGGAAACGCCTGGTTCTTCATCCCCAGCGCCATCTTACTTGGTGCG

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 751
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 1270

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K549003 short</partinfo>
-The biobrick contains the RcnA promotor from E. coli fused with the luxAB reporter gene. E coli strains own the RcnR-regulator gene which direct regulates the transcription of the RcnA nickel cobalt efflux protein from which the promotor is amplified. In presence of nickel the Luciferase is expressed.
+The biobrick contains the RcnA promotor from E. coli fused with the ''luxAB'' reporter gene. ''E. coli'' strains own the RcnR-regulator gene which directly regulates the transcription of the RcnA nickel-cobalt efflux protein from which the promotor is amplified.
+In absence of Ni(II), RcnR represses the ''rcnA''-promoter.
+After addition of Ni (II) (NiCl2), RcnR binds this ion and is released from the promoter. Therefore, luxAB will be expressed.
+--> also use luxCDE or luciferine to detect output.
+In presence of nickel the Luciferase is expressed.
 The promoter is also a little bit sensitive for Co 2+.