Difference between revisions of "Part:BBa K1412007"
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<partinfo>BBa_K1412007 short</partinfo>
 
<partinfo>BBa_K1412007 short</partinfo>
  
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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.
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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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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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=='''Usage'''==
 
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1.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 ].
 
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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.
 
  
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=='''Method'''==
  
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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 ([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 ).
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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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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.
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=='''Experimental data''' ==
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<div style="float:left">[[Image:IMG_20140825_185642.jpg|300px]];<div style="clear:both"></div>
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<partinfo>BBa_K1412007 parameters</partinfo>
 
<partinfo>BBa_K1412007 parameters</partinfo>
 
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=='''Reference'''==
 
=='''Reference'''==
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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 ].
 
[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]