Difference between revisions of "Part:BBa K1412007"
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__NOTOC__
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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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<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 <i>CheZ</i> 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 <i>CheZ</i>-/- cells. It has been reported that <i>CheZ</i>-/- strain has a higher frequency of direction change and thus a narrower range of mobility.
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As iGEM14_XMU-China has developed a system to characterize the strength of promoters and RBSs, 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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=='''Method'''==
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1.We performed a swarming assay to characterize the motility of ''<i>CheZ</i>''-deficient strain CL1 and cells transformed with ''<i>CheZ</i>''-expressing plasmid [https://parts.igem.org/Part:BBa_K1412001  BBa_K1412001 ].
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Our results clearly show that ''<i>CheZ</i>''-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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2.We also performed a swarming assay to characterize the strength of RBS with CL1 cells. Three strains of CL1 are transformed with ''<i>CheZ</i>''-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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Changing different RBS in the same circuit, we can characterize the efficiency of RBS, since the expression strength of ''<i>CheZ</i>'' 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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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1412007 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K1412007 parameters</partinfo>
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Contructing new device by this part, then characterize the device in <i>CL-1</i>(knock <i>CheZ</i> gene out of genome). In order for reference, we need to choose appropriate reference parts(see the wiki from iGEM14_XMU-China) which has only one portion(RBS or promoter) different from the new contructed part. As the expression strength from reference part is already known, we can spot both kinds of device on the same semi-solid medium culture. By comparing the chemotaxis diameter of both devices, we can tell unknown expression strength from unknown promoters or RBSs.
  
 
=='''Reference'''==
 
=='''Reference'''==

Revision as of 16:03, 9 October 2014

This part is derived from BBa_K1412005 by taking RBS out.

As iGEM14_XMU-China has developed a system to characterize the strength of promoters and RBSs, 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 CL-1(knock CheZ gene out of genome). In order for reference, we need to choose appropriate reference parts(see the wiki from iGEM14_XMU-China) which has only one portion(RBS or promoter) different from the new contructed part. As the expression strength from reference part is already known, we can spot both kinds of device on the same semi-solid medium culture. By comparing the chemotaxis diameter of both devices, we can tell unknown expression strength from unknown promoters or RBSs.

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 ].