Difference between revisions of "Part:BBa K1412007"

(Created page with "__NOTOC__ <partinfo>BBa_K1412005 short</partinfo> This part consists of a cheZ coding region which is responsible for the dephosphorylation of CheY protein in bacteria flagella ...")
 
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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K1412005 short</partinfo>
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<partinfo>BBa_K1412007 short</partinfo>
  
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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.
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This part is derived from [https://parts.igem.org/Part:BBa_K1412005 BBa_K1412005] by taking RBS out.
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This part also has a RBS coding region whose efficiency is 1.0.
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===Biology===
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As iGEM14_[http://2014.igem.org/Team:XMU-China# XMU-China] has developed a system to characterize the strength of different promoter and RBS, this part provides an easier assembly path to contruct new devices to characterize the strength of promoters and RBSs which is unknown.
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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.
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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.
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===Usage===
 
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We performed a swarming assay to characterize the motility of cheZ-deficient strain CL1 and cells transformed with cheZ-expressing plasmid.(BBa_K1412001).
 
  
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Our results clearly show that cheZ-deficient mutants show smaller diffusion, while they are rescued by our BBa_K1412001 plasmid.
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=='''Method'''==
  
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<div style="float:left">[[Image:IMG_20140825_185642.jpg|300px]];<div style="clear:both"></div>
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Contructing new device by this part, then characterize the device in <i>E.coli (CL-1)</i>(knock <i>CheZ</i> gene out of genome). In order to take reference, we need to choose appropriate reference parts(see the wiki from iGEM14_[http://2014.igem.org/Team:XMU-China# XMU-China]) which has only one portion(RBS or promoter) different from the new contructed device. As the expression strength from reference part is already known, we can spot both kinds of device on the same semi-solid culture medium. By comparing the chemotaxis diameter of both devices, we can tell unknown expression strength of unknown promoters or RBSs.
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===Reference===
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[1] 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] Chelsky D and Dahlquist FW (1980) Chemotaxis in Escherichia coli: association of protein components. Biochemistry 19: 4633–4639
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1412005 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1412007 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K1412005 parameters</partinfo>
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<partinfo>BBa_K1412007 parameters</partinfo>
 
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=='''Reference'''==
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[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 ].
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<I><B>More information, click here: [http://2014.igem.org/Team:XMU-China# XMU-China]

Latest revision as of 17:00, 14 October 2014

CheZ-TT

This part is derived from BBa_K1412005 by taking RBS out.

As iGEM14_[http://2014.igem.org/Team:XMU-China# XMU-China] has developed a system to characterize the strength of different promoter and RBS, this part provides an easier assembly path to contruct new devices to characterize the strength of promoters and RBSs which is unknown.


Method

Contructing new device by this part, then characterize the device in E.coli (CL-1)(knock CheZ gene out of genome). In order to take reference, we need to choose appropriate reference parts(see the wiki from iGEM14_[http://2014.igem.org/Team:XMU-China# XMU-China]) which has only one portion(RBS or promoter) different from the new contructed device. As the expression strength from reference part is already known, we can spot both kinds of device on the same semi-solid culture medium. By comparing the chemotaxis diameter of both devices, we can tell unknown expression strength of unknown promoters or RBSs.


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 ].

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