Difference between revisions of "Part:BBa K389010"

Revision as of 13:54, 4 October 2010

VirA receptor (expressed)

This BioBrick contains a VirA receptor from Agrobacterium tumefaciens C58 under the control of a medium strong constitutive promoter.

Usage and Biology

We used this BioBrick to mutate virA by error-prone PCR so the mutated VirA receptor is expressed and ready to be screened for several substances.

The screening system for a mutated virA gene contains this BioBrick in a plasmid with ColE1 ori and the BioBrick BBa_K389011 in a plasmid with R6K ori. Both plasmids will be transformed to and screened in E. coli EC100D. Plasmids with R6K ori are only replicated in pir+ or pir116 E. coli strains. Once a receptor with high sensitivity for a screened substance is found, the plasmids are isolated and transformed to e.g. E. coli TOP10. Because the R6K ori does not work in this strain because it is not a pir+ / pir116 strain, it is easy to separate the mutated virA BioBrick from the screening plasmid.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 647
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1632
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1768

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K389010 short</partinfo>
-This BioBrick contains a ''virA'' receptor from ''Agrobacterium tumefaciens'' C58 under the control of a constitutive promotor.
+This BioBrick contains a VirA receptor from ''Agrobacterium tumefaciens'' C58 under the control of a medium strong constitutive promoter.
-We used it to mutate ''virA'' by error-prone PCR. For screening the mutants BBa_K389011 is also needed.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+We used this BioBrick to mutate ''virA'' by error-prone PCR so the mutated VirA receptor is expressed and ready to be screened for several substances.
+The screening system for a mutated ''virA'' gene contains this BioBrick in a plasmid with ColE1 ori and the BioBrick <partinfo>K389011</partinfo> in a plasmid with R6K ori. Both plasmids will be transformed to and screened in ''E. coli'' EC100D. Plasmids with R6K ori are only replicated in pir+ or pir116 ''E. coli'' strains. Once a receptor with high sensitivity for a screened substance is found, the plasmids are isolated and transformed to ''e.g. E. coli'' TOP10. Because the R6K ori does not work in this strain because it is not a pir+ / pir116 strain, it is easy to separate the mutated ''virA'' BioBrick from the screening plasmid.
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