Difference between revisions of "Part:BBa K1412000"

Latest revision as of 23:19, 17 October 2014

CheZ generator under Lac promoter

This part consists of Lac promoter and CheZ generator. It could endow chemotaxis ability to CL-1.

Usage and Biology

CheZ can dephosphorylate CheY-P to CheY. CheY could make the flagellar motor rotate counterclockwise so that the cell could swim. Without CheZ, CheY-P is phosphorylated, which could bind to the flagellar motor protein FliM, causing the cell to tumble. The expression of CheZ allows E.coli to navigate gradients of natural chemoattractants.

At the same time, we know that Lac promoter is an inverting regulator sensitive to LacI and CAP, while LacI can be inhibited by IPTG. With LacI protein, CAP protein and IPTG, we can adjust the expression of CheZ via Lac promoter to control the movement of E.coli. The part BBa_K1412001 has already been used to govern E.coli chemotaxis successfully.

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
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Experimental Data

Figue 1. The comparison of chemotaxis ability between device BBa_J04450 and BBa_K1412000

CheZ and LacI are both knocked out of the genome of CL-1, thus CL-1 does not have the ability of chemotaxis and will not inhibit the expression of Lac promoter. Therefore CL-1 could express CheZ in BBa_K1412000 to regain chemotaxis ability. While compare with BBa_J04450, which has not the ability of chemotaxis. We can get the result shown on the figure, the diameter of chemotaxis: d2 > d1. It mean that BBa_K1412000 have the chemotaxis ability.

Protocol

We used M63 semi-solid plate to observe the behavior of the two E.coli(CL-1) strain. We spotted 3μL of the cells on the same plate, then cultured the plate at 37℃ incubator. By comparing the chemotaxis diameter of the bacteria on the plate, we conclude that BBa_K1412000 could endow chemotaxis ability to CL-1.

References

[1] [http://pubs.acs.org/doi/abs/10.1021/ja0692480 Topp S, Gallivan J P. Guiding bacteria with small molecules and RNA[J]. Journal of the American Chemical Society, 2007, 129(21): 6807-6811.].

More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]

@@ Line 5: / Line 5: @@
-This part is consist of Lac promoter and CheZ generator.
+This part consists of Lac promoter and CheZ generator. It could endow chemotaxis ability to <i>CL-1</i>.
-CheZ can dephosphorylate CheY to make the flagellar motor rotate counterclockwise so that the cell to run and tumble.Without CheZ, CheY is phosphorylated, which can bind to the flagellar motor protein FliM, causing the cell to tumble.so the cell tumbles only.
+=='''Usage and Biology'''==
+CheZ can dephosphorylate CheY-P to CheY. CheY could make the flagellar motor rotate counterclockwise so that the cell could swim. Without CheZ, CheY-P is phosphorylated, which could bind to the flagellar motor protein FliM, causing the cell to tumble.
 The expression of CheZ allows <i>E.coli</i> to navigate gradients of natural chemoattractants.
-At the same time, we know that Lac promoter is an inverting regulator sensitive to LacI and CAP, while LacI can be inhibited by IPTG. So with LacI protein, CAP protein and IPTG,we can adjust the expression of CheZ via Lac promoter to control the movement of <i>E.coli</i>.
+At the same time, we know that Lac promoter is an inverting regulator sensitive to LacI and CAP, while LacI can be inhibited by IPTG. With LacI protein, CAP protein and IPTG, we can adjust the expression of CheZ via Lac promoter to control the movement of <i>E.coli</i>.
-[[File:pLac-RBS-CheZ-TT.png]]
+The part [https://parts.igem.org/Part:BBa_K1412001 BBa_K1412001] has already been used to govern <i>E.coli</i> chemotaxis successfully.
-==='''Usage and Biology'''===
 ----
@@ Line 23: / Line 26: @@
 <partinfo>BBa_K1412000 SequenceAndFeatures</partinfo>
-==='''Experimental Data'''===
+=='''Experimental Data'''==
 [[File:CL-1 and BBa K1412000.png |500px]]
-Figure 2. Comparision with [https://parts.igem.org/Part:BBa_J04450 BBa_J04450] and BBa_K1412000
+'''Figue 1. The comparison of chemotaxis ability between device <bbpart>BBa_J04450</bbpart> and <bbpart>BBa_K1412000</bbpart>'''
-BBa_J04450 and BBa_K1412000 are transformed into <i>CL-1</i> strains respectively. From the result, we can see that the chemotaxis diameter of BBa_K1412000 (d2) is longer than BBa_J04450 (d1), which means BBa_K1412000 could work as expectation.
+<i>CheZ</i> and <i>LacI</i> are both knocked out of the genome of <i>CL-1</i>, thus <i>CL-1</i> does not have the ability of chemotaxis and will not inhibit the expression of Lac promoter. Therefore <i>CL-1</i> could express <i>CheZ</i> in <bbpart>BBa_K1412000</bbpart> to regain chemotaxis ability. While compare with <bbpart>BBa_J04450</bbpart>, which has not the ability of chemotaxis. We can get the result shown on the figure, the diameter of chemotaxis: d2 > d1. It mean that <bbpart>BBa_K1412000</bbpart> have the chemotaxis ability.
+==='''Protocol'''===
+----
-We use M63 semi-solid plate to observe the behavior of the two <i>E.coli(CL-1)</i> strain. We dot 3μL of the two bacterias on the same plate, then place the plate at 37℃. By comparing the chemotaxis diameter of the bacteria on the plate, we conclude BBa_K1412000 has the ability of chemotaxis.
+We used M63 semi-solid plate to observe the behavior of the two <i>E.coli(CL-1)</i> strain. We spotted 3μL of the cells on the same plate, then cultured the plate at 37℃ incubator. By comparing the chemotaxis diameter of the bacteria on the plate, we conclude that BBa_K1412000 could endow chemotaxis ability to <i>CL-1</i>.
+===<B>References</B>===
+[1] [http://pubs.acs.org/doi/abs/10.1021/ja0692480 Topp S, Gallivan J P. Guiding bacteria with small molecules and RNA[J]. Journal of the American Chemical Society, 2007, 129(21): 6807-6811.].
 ----
+<I><B>More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]
 <!-- Uncomment this to enable Functional Parameter display
 ===Functional Parameters===
 <partinfo>BBa_K1412000 parameters</partinfo>
 <!-- -->