Difference between revisions of "Part:BBa I712019:Experience"

Revision as of 01:18, 2 November 2017

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_I712019

User Reviews

UNIQ60bfb0da62063694-partinfo-00000000-QINU

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theos

The 2011 SJTU-BioX-iGEM team has used this part to further construct vectors including BBa_K567003-BBa_K567010. Functional analysis of these luciferases were performed with luciferin reaction. This part has been great!

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theos

The Cambridge team confirmed that this part did emit light from E. coli when placed under an expression vector (:BBa_J13002) with the addition of D-luciferin. Despite the claim that the "part contains a Eukaryotic ribosome binding site and non-standard prefix and suffix" we were able to express it in E. coli after adding a prokaryotic RBS and promoter using standard BioBrick assembly.

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Catramp

The Utah_State 2011 iGEM Team has confirmed that this part emits light when placed under control of a promoter and D-luciferin is added, and is capable of working with standard luciferase testing kits and luminometers.

Improvements: This part has been improved upon with the addition of Dual Luciferase Assay intermediates that make construction of promoter testing devices with this part much easier (Dual Luciferase Assay Component).

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UT-Tokyo

UT-Tokyo 2011 Team has characterized that this part emits luminescence with a promoter and substrate (D-luciferin). For testing this device, we carried out a promoter assay using BBa_R0011 and BBa_J23119 as a control. We succesfully evaluated the relative expression levels of BBa_R0011 with various IPTG concentration, compared to that of BBa_J23119. For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page].

Improvements: UT-Tokyo 2011 team has utilized this part to devise a Firefly-Renilla Dual Luciferase Assay Kit. This kit makes an evaluation of promoters and other cis-elements much easyer and more quantitative than existing devices (including GFP-reporter system). We also conducted a calibration of Photinus pyralis luciferase using a standard reagent, demonstrating that intracellular luciferase amount can be directly estimated from relative luminescence unit (RPU) by a linear regression. For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page].

Improvements:

This part was improved by NUDT_CHINA in iGEM 2017 to optimize its function in a dual-luciferase miRNA target expression system. Two restriction sites of BsmBI were added to the 3’ end of firefly luciferase, through which, the 3’ untranslated region of any gene can be inserted using Golden-Gate Assembly, with no worry about the multiple illegal sites contained in it. For more information, please visit our part page: https://parts.igem.org/Part:BBa_K2440008.

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UNIQ60bfb0da62063694-partinfo-00000005-QINU The Tianjin 2010 igem team confirmed that this part works well since we have used this part to construct our parts which separate the luciferase gene. Our sequencing result reveal that the quality of this part is ok. UNIQ60bfb0da62063694-partinfo-00000006-QINU

@@ Line 50: / Line 50: @@
 UT-Tokyo 2011 team has utilized this part to devise a [https://parts.igem.org/Part:BBa_K518002 Firefly-Renilla Dual Luciferase Assay Kit]. This kit makes an evaluation of promoters and other cis-elements much easyer and more quantitative than existing devices (including GFP-reporter system).
 We also conducted a calibration of Photinus pyralis luciferase using a standard reagent, demonstrating that intracellular luciferase amount can be directly estimated from relative luminescence unit (RPU) by a linear regression. For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page].
+'''Improvements:'''
+This part was improved by NUDT_CHINA in iGEM 2017 to optimize its function in a dual-luciferase miRNA target expression system. Two restriction sites of BsmBI were added to the 3’ end of firefly luciferase, through which, the 3’ untranslated region of any gene can be inserted using Golden-Gate Assembly, with no worry about the multiple illegal sites contained in it. For more information, please visit our part page: https://parts.igem.org/Part:BBa_K2440008.
 |};