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. 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> | + | 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. <\n>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> |
<p><img src="https://static.igem.org/mediawiki/2018/4/42/T--SHSBNU_China--Parts_Regis.png" style="width:50%"/></image></p> | <p><img src="https://static.igem.org/mediawiki/2018/4/42/T--SHSBNU_China--Parts_Regis.png" style="width:50%"/></image></p> | ||
<p class="text"> | <p class="text"> |
Revision as of 20:24, 17 October 2018
CotA Laccase of B.subtilis
CotA Laccase is an endospore type protein secreted from B.subtilis
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1348
- 1000INCOMPATIBLE 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. <\n>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.