Difference between revisions of "Part:BBa K2684000"
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<h2>Overview</h2>
 
<h2>Overview</h2>
 
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Part BBa_K2684000 was improved from previous part <a href="https://parts.igem.org/Part:BBa_K1336002">BBa_K1336002</a>. Our team made a point mutation to eliminate the EcoRI cutting site so that we can meet the standard of RFC10, a commonly-used standard for interchangeable parts which are based on idempotent assembly. <\br>We tested the enzyme activity of our part based on the weight of our sample. Formula: laccase activity (nmol/min/g) = ΔA /ε(ABTS mmolar extinction coefficient) / d * V (total volume of reaction) / V (volume of sample in the reaction, 0.025mL) / W (sample mass, g) * V (extracted liquid added, 1mL) / T (reaction time, 3 minutes) = 130 *ΔA / W. The result is shown as following: </p>
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Part BBa_K2684000 was improved from previous part <a href="https://parts.igem.org/Part:BBa_K1336002">BBa_K1336002</a>. Our team made a point mutation to eliminate the EcoRI cutting site so that we can meet the standard of RFC10, a commonly-used standard for interchangeable parts which are based on idempotent assembly. <br>We tested the enzyme activity of our part based on the weight of our sample. Formula: laccase activity (nmol/min/g) = ΔA /ε(ABTS mmolar extinction coefficient) / d * V (total volume of reaction) / V (volume of sample in the reaction, 0.025mL) / W (sample mass, g) * V (extracted liquid added, 1mL) / T (reaction time, 3 minutes) = 130 *ΔA / W. The result is shown as following: </p>
 
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<p><img src="https://static.igem.org/mediawiki/2018/4/42/T--SHSBNU_China--Parts_Regis.png" style="width:50%"/></image></p>
 
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Revision as of 20:25, 17 October 2018


CotA Laccase of B.subtilis

CotA Laccase is an endospore type protein secreted from B.subtilis

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1348
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 286

Overview

Part BBa_K2684000 was improved from previous part BBa_K1336002. Our team made a point mutation to eliminate the EcoRI cutting site so that we can meet the standard of RFC10, a commonly-used standard for interchangeable parts which are based on idempotent assembly.
We tested the enzyme activity of our part based on the weight of our sample. Formula: laccase activity (nmol/min/g) = ΔA /ε(ABTS mmolar extinction coefficient) / d * V (total volume of reaction) / V (volume of sample in the reaction, 0.025mL) / W (sample mass, g) * V (extracted liquid added, 1mL) / T (reaction time, 3 minutes) = 130 *ΔA / W. The result is shown as following:

The improvement is valid since it was normally produced.

References

Guan, Z.-B., Luo, Q., Wang, H.-R., Chen, Y., & Liao, X.-R. (2018). Bacterial laccases: promising biological green tools for industrial applications. Cellular and Molecular Life Sciences.

“Help:Standards/Assembly/RFC10.” Help:Standards/Assembly/RFC10, parts.igem.org/Help:Standards/Assembly/RFC10.