Difference between revisions of "Part:BBa K283002"

Latest revision as of 12:10, 21 October 2009

Speed control device (pLacI-rbs-TetR-terminator-ptet-rbs-chez_his-double terminators)

To test if cheZ expression level was proportional to the bacterial swimming speed and hopefuly to control the speed, we designed and made this speed controller device. The full length cheZ gene fused with His6-tag was put under the control of pTet, whose expression is repressed by constitutively expressed TetR and can be induced by the addition of anhydrotetracycline or tetracycline. Since lacking antibody to recognize the CheZ protein, His6-tag was added to facilitate the measurement. This device has been demonstrated to work as expected in our system (see experience or visit our wiki).This part does not pose any biological threat.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1782
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K283002 short</partinfo>
-This is a device that enable the control of chez(histag) using anhydrotetracycline or tetracycline. The pLacI promoter(R0010)enables high level of expression of the TetR repressor(C0040) protein.Without the presence of LacI repressor molecule, the pLacI promoter is constitutively on to express the TetR repressor protein.Therefore the ptet(R0040) promoter is repressed under normal condition. However, in the presence of anhydrotetracycline or tetracycline, which binds to TetR and remove its inhibiting effect on pTet promoter, the chez-histag(K283008) protein can be expressed. Consequently, the expression level of chez-histag,which responsible for the dephosphorylation of cheY protein in bacteria flagella movement, should be controlled by the concentration of anhydrotetracycline or tetracycline.
+To test if ''cheZ'' expression level was proportional to the bacterial swimming speed and hopefuly to control the speed, we designed and made this speed controller device. The full length ''cheZ'' gene fused with His6-tag was put under the control of pTet, whose expression is repressed by constitutively expressed TetR and can be induced by the addition of anhydrotetracycline or tetracycline. Since lacking antibody to recognize the CheZ protein, His6-tag was added to facilitate the measurement. This device has been demonstrated to work as expected in our system (see experience or visit our wiki).This part does not pose any biological threat.
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