Difference between revisions of "Part:BBa J10050:Experience"
 
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===Applications of BBa_J10050===
 
===Applications of BBa_J10050===
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Lambert_GA 2019 used this part in experiments to characterize the effect of different strengths of promoters and ribosomal binding sites on expression. The data obtained can aid in the design of constructs by giving information about controlling expression.
  
 
===User Reviews===
 
===User Reviews===
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<partinfo>BBa_R0071 AddReview 4</partinfo>
 
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<I>ETH Zurich 2014</I>
 
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= Characterization of weak promoter (BBa_J23113) and weak RBS (BBa_B0031) =
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= Lambert_GA 2019 Characterization =
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==Characterization of weak promoter (BBa_J23113) and weak RBS (BBa_B0031)==
  
 
Differences in promoter strength had more effect on enzymatic expression compared to differences in RBS strengths as evidenced in the graphs below. Lambert_GA 2019 had originally thought that BBa_J10050 would result in the least amount of expression out of the tested combinations, but this part, along with BBa_J10051 and BBa_J10052, all shared the lowest expression out of the combinations. This is due to the translation rate not being an important factor in the determination of expression of ONP; however, the promoter strength is a significant factor. In other words, expression values are significantly different with different promoters but not with different RBS strengths.
 
Differences in promoter strength had more effect on enzymatic expression compared to differences in RBS strengths as evidenced in the graphs below. Lambert_GA 2019 had originally thought that BBa_J10050 would result in the least amount of expression out of the tested combinations, but this part, along with BBa_J10051 and BBa_J10052, all shared the lowest expression out of the combinations. This is due to the translation rate not being an important factor in the determination of expression of ONP; however, the promoter strength is a significant factor. In other words, expression values are significantly different with different promoters but not with different RBS strengths.
  
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[[File:ETH Zurich 1crosstalkPrhl.png|400px|thumb|center| '''Figure 1 Overview of possible crosstalk of the [https://parts.igem.org/Part:BBa_C0171 RhlR]/[https://parts.igem.org/Part:BBa_I14017 pRhl] system with three different [[AHL|AHLs]].''' Usually, [[AHL|C4-HSL]] binds to its corresponding regulator, [https://parts.igem.org/Part:BBa_C0171 RhlR], and activates the [https://parts.igem.org/Part:BBa_I14017 pRhl] promoter (green). However, [https://parts.igem.org/Part:BBa_C0171 RhlR] may also bind [[AHL|3OC12-HSL]] (red) or [[3OC6HSL|3OC6-HSL]] (light blue) and then unintentionally activate [https://parts.igem.org/Part:BBa_I14017 pRhl].]]
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<center>[[File:T--Lambert GA--tuning1a.png|800px]]</center>
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<center><i>Figure 1: This sequence is to be used as a proof of concept, and can be used to develop biosensor control devices. The highlighted points in yellow represent BBa_J10050 (weak promoter/ weak RBS) compared to BBa_J10051 (weak promoter/ strong RBS) and BBa_J10052 (weak promoter/ medium RBS), both in light blue. The weak, medium, and strong RBS have similar expression levels when in combination with the weak promoter.</i></center>
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<center>[[File:T--Lambert GA--tuning1b.png|800px]]</center>
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<center><i>Figure 2: The highlighted points represent the BBa_J10050 composite part (weak promoter/ weak RBS). As the promoter strength increases, the expression increases as well.</i>
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The graphs above show that RBS strength has a negligible effect on expression, whereas increased promoter strength increases expression. </center>
  
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[[File:ETH_Zurich_2crosstalkPrhl.png|thumb|400px|center|'''Figure 2 Overview of possible crosstalk of the [https://parts.igem.org/Part:BBa_C0171 RhlR]/[https://parts.igem.org/Part:BBa_i14017 pRhl] system with two additional regulators ([https://parts.igem.org/Part:BBa_R0062 LuxR] and [https://parts.igem.org/Part:BBa_C0179 LasR]).''' Usually, [https://parts.igem.org/Part:BBa_C0171 RhlR] together with inducer [[AHL|C4-HSL]] activate their corresponding promoter [https://parts.igem.org/Part:BBa_I14017 pRhl] (green). However, [https://parts.igem.org/Part:BBa_I14017 pRhl] may also be activated by the [https://parts.igem.org/Part:BBa_C0062 LuxR] regulator together with [[3OC6HSL|3OC6-HSL]] (light blue) or by the [https://parts.igem.org/Part:BBa_C0179 LasR] regulator together with [[AHL|3OC12-HSL]] (red).]]
 
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[[File:ETH Zurich 2014 2nd order rhl.png|400px|thumb|center| '''Figure 3 Overview of possible crosstalk of the [https://parts.igem.org/Part:BBa_I14017 pRhl] promoter with both the regulator and inducer being unrelated to the promoter and each other.''' Usually, [https://parts.igem.org/Part:BBa_C0171 RhlR] together with inducer [[AHL|C4-HSL]] activate their corresponding promoter [https://parts.igem.org/Part:BBa_I14017 pRhl] (green). However, [https://parts.igem.org/Part:BBa_I14017 pRhl] may also be activated by another regulator together with an unrelated inducer.  For example, the inducer [[3OC6HSL|3OC6-HSL]] (light blue) may interact with the [https://parts.igem.org/Part:BBa_C0179 LasR] regulator (red) and together activate [https://parts.igem.org/Part:BBa_I14017 pRhl] (green).]]
 
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<partinfo>BBa_I14017 AddReview 0</partinfo>
 
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<I>Antiquity</I>
 
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This review comes from the old result system and indicates that this part did not work in some test.
 
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Latest revision as of 19:52, 21 October 2019


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_J10050

Lambert_GA 2019 used this part in experiments to characterize the effect of different strengths of promoters and ribosomal binding sites on expression. The data obtained can aid in the design of constructs by giving information about controlling expression.

User Reviews

UNIQ578a76558c394b93-partinfo-00000000-QINU

Lambert_GA 2019 Characterization

Characterization of weak promoter (BBa_J23113) and weak RBS (BBa_B0031)

Differences in promoter strength had more effect on enzymatic expression compared to differences in RBS strengths as evidenced in the graphs below. Lambert_GA 2019 had originally thought that BBa_J10050 would result in the least amount of expression out of the tested combinations, but this part, along with BBa_J10051 and BBa_J10052, all shared the lowest expression out of the combinations. This is due to the translation rate not being an important factor in the determination of expression of ONP; however, the promoter strength is a significant factor. In other words, expression values are significantly different with different promoters but not with different RBS strengths.


T--Lambert GA--tuning1a.png


Figure 1: This sequence is to be used as a proof of concept, and can be used to develop biosensor control devices. The highlighted points in yellow represent BBa_J10050 (weak promoter/ weak RBS) compared to BBa_J10051 (weak promoter/ strong RBS) and BBa_J10052 (weak promoter/ medium RBS), both in light blue. The weak, medium, and strong RBS have similar expression levels when in combination with the weak promoter.



T--Lambert GA--tuning1b.png


Figure 2: The highlighted points represent the BBa_J10050 composite part (weak promoter/ weak RBS). As the promoter strength increases, the expression increases as well.



The graphs above show that RBS strength has a negligible effect on expression, whereas increased promoter strength increases expression.













UNIQ578a76558c394b93-partinfo-00000001-QINU