Difference between revisions of "Part:BBa K1412005"
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<partinfo>BBa_K1412005 short</partinfo> | <partinfo>BBa_K1412005 short</partinfo> | ||
| − | This part consists of a CheZ coding region which is responsible for the dephosphorylation of CheY protein in | + | This part consists of a CheZ coding region which is responsible for the dephosphorylation of CheY-P protein in <i>E.coli</i> chemotaxis system. This part does not pose any biological threat. With a promoter of you interest, this device can endow chemotaxis ability to<i>.E.coli CL-1</i> whose <i>CheZ</i> gene is knocked out of genome. It has been reported that ΔCheZ strain has a higher frequency of direction change and thus resulting low chemotaxis ability with a narrower range of mobility. |
This part also has a RBS coding region whose efficiency is 1.0. | This part also has a RBS coding region whose efficiency is 1.0. | ||
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=='''Biology''' == | =='''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. | + | Chemotaxis in <i>E.coli</i> 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-P 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 the movement performed by bacteria towards an attractant or away from a repellent. It 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 | + | 1. We performed a swarming assay to characterize the motility of ''CheZ''-deficient strain <I>CL-1</I> and cells transformed with ''CheZ''-expressing plasmid [https://parts.igem.org/Part:BBa_K1412001 BBa_K1412001]. |
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| + | 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. | ||
| − | + | 2. We also performed a swarming assay to characterize the strength of RBS with <I>CL-1</I> cells. Three strains of <i>CL-1</i> 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). | |
| + | 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''' == | =='''Experimental data''' == | ||
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=='''Reference'''== | =='''Reference'''== | ||
| − | [1] Paungfoo-Lonhienne C et al. (2010) Turning the table: plants consume microbes as a source of nutrients. PLoS One 5(7): e11915. | + | [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]. |
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| + | [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]. | ||
| + | ---- | ||
| − | [ | + | <I><B>More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China] |
Latest revision as of 17:56, 14 October 2014
RBS (1.0) CheZ double terminators
This part consists of a CheZ coding region which is responsible for the dephosphorylation of CheY-P protein in E.coli chemotaxis system. This part does not pose any biological threat. With a promoter of you interest, this device can endow chemotaxis ability to.E.coli CL-1 whose CheZ gene is knocked out of genome. It has been reported that ΔCheZ strain has a higher frequency of direction change and thus resulting low chemotaxis ability with a narrower range of mobility. This part also has a RBS coding region whose efficiency is 1.0.
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-P 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. It is controlled by a number of chemotaxis proteins that make up a signalling pathway, please see the demonstration below for details.
Usage
1. We performed a swarming assay to characterize the motility of CheZ-deficient strain CL-1 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 the strength of RBS with CL-1 cells. Three strains of CL-1 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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].
More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]