Difference between revisions of "Part:BBa K2717023:Experience"

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===Applications of BBa_K2717023===
 
===Applications of BBa_K2717023===
  
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We used In-Fusion Cloning tech to link our composite to pUCyder. The whole pathway can be described in Figure 1.  
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We used In-Fusion Cloning tech to link our composite to pUCyder.  
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Figure 1 the constuction of part BBa_K2717018
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Then the plasmid is transfered into E.coli BL21. We use Flow Cytometer (FCM) to mearsure the bacteria's number and a relative fluorescence. Figure 2 is one of the FCM result graphs. The specific experiments and data processing methods can be found in our wiki. Finally, by comparing the final relative fluorescent intensity with that of control group, we found the exogenous CI sequence causes a 150% improvement to pink fluorescent intensity. That is to say, CI protein can indeed increase the expression of following gene.
 
Then the plasmid is transfered into E.coli BL21. We use Flow Cytometer (FCM) to mearsure the bacteria's number and a relative fluorescence. Figure 2 is one of the FCM result graphs. The specific experiments and data processing methods can be found in our wiki. Finally, by comparing the final relative fluorescent intensity with that of control group, we found the exogenous CI sequence causes a 150% improvement to pink fluorescent intensity. That is to say, CI protein can indeed increase the expression of following gene.
  
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Table 1 the comparison of GFP expression between CI-activation and control.
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https://static.igem.org/mediawiki/2018/6/65/T--BNU-China--_R18.png
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Fig.18 A) The western blot image of CI, marker on the right. The protein is between 34kD and 25kD, which is precisely the exact molecular weight of itself. B) Data of the induction test. Each row indicates one time point and the columns from left to right represent IEtot of experiment strain, IEtot of the control strain and CI effect respectively. C) Bar gram drawn using the data presented in c to give a more intuitive impression.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 21:48, 17 October 2018


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K2717023

We used In-Fusion Cloning tech to link our composite to pUCyder.


Then the plasmid is transfered into E.coli BL21. We use Flow Cytometer (FCM) to mearsure the bacteria's number and a relative fluorescence. Figure 2 is one of the FCM result graphs. The specific experiments and data processing methods can be found in our wiki. Finally, by comparing the final relative fluorescent intensity with that of control group, we found the exogenous CI sequence causes a 150% improvement to pink fluorescent intensity. That is to say, CI protein can indeed increase the expression of following gene.


T--BNU-China--_R18.png


Fig.18 A) The western blot image of CI, marker on the right. The protein is between 34kD and 25kD, which is precisely the exact molecular weight of itself. B) Data of the induction test. Each row indicates one time point and the columns from left to right represent IEtot of experiment strain, IEtot of the control strain and CI effect respectively. C) Bar gram drawn using the data presented in c to give a more intuitive impression.

User Reviews

UNIQb7749b09c40c2878-partinfo-00000000-QINU UNIQb7749b09c40c2878-partinfo-00000001-QINU