Difference between revisions of "Part:BBa K313008"

Latest revision as of 19:34, 5 November 2010

pT7 - Cre recombinase generator

It is the generator of the Cre recombinase.

Cre is transcribed by the T7 RNA polymerase.

By this construct, we conducted recombination experiment along with "Cre recombinase assay device". (Please see here.)

This construct also serves as a positive control of "Cre generator with terminator" series as shown the table below.

Other Cre generator with terminator parts
number	description	Subparts compositon	PartsID
1	Cre generator with single terminator	pT7-t90%-rbs-cre-dt	BBa_K313017
2	Cre generator with double terminator	pT7-t90%-t90%-rbs-cre-dt	BBa_K313018
3	Cre generator with weak single terminator	pT7-t80%-rbs-cre-dt	BBa_K313021
4	Cre generator with weak double terminator	pT7-t80%-t80%-rbs-cre-dt	BBa_K313022

Now we are planning to conduct further research, seeking for appropriate terminator for 4C3 leak-switch.

Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw Terminator leak] assay page and our [http://2010.igem.org/Team:UT-Tokyo/Sudoku_experiments#Terminator_leak wiki].

verification experiments

This part was ligated with loxP-gfp-loxP sequence and transformed into E.coli. By incubating E.coli, then extracting plasmids, and sequencing them, it was proved that gfp sequence flanked by loxP sequence was removed. As a conclusion, cre generated by BBa_K313008 is able to catalyze site specific recombination reaction.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 431
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 117
1000
COMPATIBLE WITH RFC[1000]

@@ Line 35: / Line 35: @@
 ===verification experiments===
-This part is ligated with loxP-gfp-loxP sequence and transformed into E.coli. By incubating E.coli, then extracting plasmids, and sequencing them, it was proved that cre generated by BBa_K313008 is able to catalyze site specific recombination reaction.
+This part was ligated with loxP-gfp-loxP sequence and transformed into E.coli. By incubating E.coli, then extracting plasmids, and sequencing them, it was proved that gfp sequence flanked by loxP sequence was removed. As a conclusion, cre generated by BBa_K313008 is able to catalyze site specific recombination reaction.
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