Difference between revisions of "Part:BBa K1412007"

Revision as of 14:25, 12 September 2014

CheZ-TT

This part is a derivative of BBa_K1412005 but weaker than it. It consists of a CheZ coding region and a RBS coding region whose relative efficiency is 0.01. This part does not pose any biological threat. With a promoter of your interest, this device rescues the mobility of CheZ-/- cells. It has been reported that CheZ-/- strain has a higher frequency of direction change and thus a narrower range of mobility.

Usage

1.We performed a swarming assay to characterize the motility of CheZ-deficient strain CL1 and cells transformed with CheZ-expressing plasmid BBa_K1412001 .

Our results clearly show that CheZ-deficient mutants show smaller diffusion, while they are rescued by our BBa_K1412001 plasmid.

2.We also performed a swarming assay to characterize characterize the strength of RBS with CL1 cells. Three strains of CL1 are transformed with CheZ-expressing plasmid of different RBS efficiency (BBa_B0034 :1.0 ; BBa_B0032 :0.3 ; BBa_B0033 :0.01 ).

Changing different RBS in the same circuit, we can characterize the efficiency of RBS, since the expression strength of CheZ is positively associated with the efficiency of RBS.

Experimental data

;

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]

Reference

[1] [http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0011915#pone-0011915-g006 Paungfoo-Lonhienne C et al. (2010) Turning the table: plants consume microbes as a source of nutrients. PLoS One 5(7): e11915 ].

[2] [http://pubs.acs.org/doi/abs/10.1021/bi00561a015 Chelsky D and Dahlquist FW (1980) Chemotaxis in Escherichia coli: association of protein components. Biochemistry 19: 4633–4639 ].

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1412007 short</partinfo>
-This part consists of a cheZ coding region which is responsible for the dephosphorylation of CheY protein in bacteria flagella movement. This part does not pose any biological threat. With a promoter of your interest, this device rescues the mobility of cheZ-/- cells. It has been reported that cheZ-/- strain has a higher frequency of direction change and thus a narrower range of mobility.
+This part is a derivative of [https://parts.igem.org/Part:BBa_K1412005   BBa_K1412005 ] but weaker than it. It consists of a CheZ coding region and a RBS coding region whose relative efficiency is 0.01. This part does not pose any biological threat. With a promoter of your interest, this device rescues the mobility of CheZ-/- cells. It has been reported that CheZ-/- strain has a higher frequency of direction change and thus a narrower range of mobility.
-This part also has a RBS coding region whose efficiency is 0.01.
-===Biology===
-Chemotaxis in E. coli is well documented. These bacteria can perform two types of movement, tumbling and smooth swimming. The difference between the two is determined by flagellar movement. During tumbling movement, the flagella move clockwise. This is caused by the formation of a complex between CheY-P and FliM, one of the flagella-associated proteins. During smooth swimming, the flagella move counter-clockwise. CheY is not phosphorylated and therefore cannot associate with flagellar proteins, causing the flagella to rotate in the opposite direction.
-Smooth swimming is the movement performed by bacteria towards an attractant or away from a repellent. Smooth swimming is controlled by a number of chemotaxis proteins that make up a signalling pathway, please see the demonstration below for details.
-===Usage===
+=='''Usage'''==
-We performed a swarming assay to characterize the motility of cheZ-deficient strain CL1 and cells transformed with cheZ-expressing plasmid.(BBa_K1412001).
+.We performed a swarming assay to characterize the motility of CheZ-deficient strain CL1 and cells transformed with CheZ-expressing plasmid [https://parts.igem.org/Part:BBa_K1412001  BBa_K1412001 ].
+Our results clearly show that CheZ-deficient mutants show smaller diffusion, while they are rescued by our [https://parts.igem.org/Part:BBa_K1412001  BBa_K1412001] plasmid.
-Our results clearly show that cheZ-deficient mutants show smaller diffusion, while they are rescued by our BBa_K1412001 plasmid.
-<div style="float:left">[[Image:IMG_20140825_185642.jpg|300px]];<div style="clear:both"></div>
+.We also performed a swarming assay to characterize characterize the strength of RBS with CL1 cells. Three strains of CL1 are transformed with CheZ-expressing plasmid of different RBS efficiency ([https://parts.igem.org/Part:BBa_B0034  BBa_B0034 ]:1.0 ; [https://parts.igem.org/Part:BBa_B0032  BBa_B0032 ]:0.3 ; [https://parts.igem.org/Part:BBa_B0033  BBa_B0033 ]:0.01 ).
-===Reference===
+Changing different RBS in the same circuit, we can characterize the efficiency of RBS, since the expression strength of CheZ is positively associated with the efficiency of RBS.
-[1] Paungfoo-Lonhienne C et al. (2010) Turning the table: plants consume microbes as a source of nutrients. PLoS One 5(7): e11915.
+=='''Experimental data''' ==
+<div style="float:left">[[Image:IMG_20140825_185642.jpg|300px]];<div style="clear:both"></div>
-[2] Chelsky D and Dahlquist FW (1980) Chemotaxis in Escherichia coli: association of protein components. Biochemistry 19: 4633–4639
@@ Line 31: / Line 31: @@
 <partinfo>BBa_K1412007 parameters</partinfo>
 <!-- -->
+=='''Reference'''==
+[1] [http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0011915#pone-0011915-g006  Paungfoo-Lonhienne C et al. (2010) Turning the table: plants consume microbes as a source of nutrients. PLoS One 5(7): e11915 ].
+[2] [http://pubs.acs.org/doi/abs/10.1021/bi00561a015  Chelsky D and Dahlquist FW (1980) Chemotaxis in Escherichia coli: association of protein components. Biochemistry 19: 4633–4639 ].