Team:Colombia Uniandes/Parts

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This is a template page. READ THESE INSTRUCTIONS.
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<h2><center>Parts</center></h2>
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You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples <a href="https://2008.igem.org/Help:Template/Examples">HERE</a>.
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Glucocorticoid sensor
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==Glucocorticoid sensor==
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Here are the parts sent to the registry:
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
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[[File:partsGC.jpg|700px|center|]]
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!align="center"|[[Team:Colombia_Uniandes|Home]]
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!align="center"|[[Team:Colombia_Uniandes/Team|Team]]
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!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Colombia_Uniandes Official Team Profile]
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{| border="1" align="center"
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!align="center"|[[Team:Colombia_Uniandes/Project|Project]]
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|+'''Name table'''
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!align="center"|[[Team:Colombia_Uniandes/Parts|Parts Submitted to the Registry]]
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| style="text-align: center;" |Part Name
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!align="center"|[[Team:Colombia_Uniandes/Modeling|Modeling]]
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| style="text-align: center;" |Registry Number
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!align="center"|[[Team:Colombia_Uniandes/Notebook|Notebook]]
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|-
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!align="center"|[[Team:Colombia_Uniandes/Safety|Safety]]
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| style="text-align: center;" |E1
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!align="center"|[[Team:Colombia_Uniandes/Attributions|Attributions]]
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| style="text-align: center;" |BBa_K1144001
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|-
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| style="text-align: center;" |E2
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| style="text-align: center;" |BBa_K1144002
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|-
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| style="text-align: center;" |E3
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| style="text-align: center;" |BBa_K1144003
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|-
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| style="text-align: center;" |E4
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| style="text-align: center;" |BBa_K1144004
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|-
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| style="text-align: center;" |E1GF
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| style="text-align: center;" |BBa_K1144005
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|-
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| style="text-align: center;" |E2GF
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| style="text-align: center;" |BBa_K1144006
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|-
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| style="text-align: center;" |E3GF
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| style="text-align: center;" |BBa_K1144007
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|-
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| style="text-align: center;" |E4GF
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| style="text-align: center;" |BBa_K1144008
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You can see the confirmation gels of the parts below:
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An important aspect of the iGEM competition is the use and creation of standard biological parts. Each team will make new parts during iGEM and will place them in the [http://partsregistry.org Registry of Standard Biological Parts]. The iGEM software provides an easy way to present the parts your team has created . The "groupparts" tag will generate a table with all of the parts that your team adds to your team sandbox.  Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.
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[[File:Gel1GC.jpg|410px|thumb|left|''' PCR confirmation: E1GF-E1GF(2)-E2GF-E2GF(2)-WM-E3GF-E3GF(2)-E4GF-E4GF(2)''']] [[File:Gel2GC.jpg|400px|thumb|center|'''Co-transformation: WM-E1GF2-E2GF-E3GF1-E3GF2-E4GF-WM- E3GF3-E4GF2-E2GF2
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''']]
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[[File:GEL3.jpg|400px|thumb|center|'''PCR confirmation of the parts cloned in the pSB1C3 backbone using the specifically designed primers 15 and 31 (see primers)''']]
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Remember that the goal of proper part documentation is to describe and define a part such that it can be used without a need to refer to the primary literature. The next iGEM team should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for  users who wish to know more.
 
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==='''Characterization of reporters'''===
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<groupparts>iGEM013 Colombia_Uniandes</groupparts>
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To assess the strength of our promoters when induced with Dexamethasone, we performed a fluorometric assay using mCherry as our reporter.Since we don't have our transactivating protein ready yet, we co-transformed our parts into the ''E. coli'' DH10B strain with the  pAT7002 vector (Aoyama and Chua, 1997), which contains a well characterized Glucocorticoid Responsive Element that also uses the GAL4 DNA binding domain.
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[[File:Response1hr.jpg|700px|thumb|center|''' Unsaturated curve where we see how the mCherry reporter appears after the addition of 10 uM dexamethasone, a glucocorticoid. Emmision intensity was measured at 607nm after excitation at 586nm''']]
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[[File:Response3hr.jpg|700px|thumb|center|''' Saturated curve for fluorescence. After three hours of the addition of 10 uM dexamethasone the reporter (mCherry) reaches its highest signal. We can see the different strengths depending on the number of UAS boxes. Emmision intensity was measured at 607nm after excitation at 586nm''']]
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We also decided to visually inspect our induced transformants. Here two of the images taken using a epifluorescent microscopy with a TRITC filter.
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[[File:E2GF.jpg|370px|thumb|left|''' Cells with the E2GF part (BBa_K1144006) showing their fluorescent reporter, mCherry, after induction with 10 uM dexamethasone! The filter used was TRITC''']]
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[[File:E4GF.jpg|370px|thumb|right|''' Cells with the E4GF part (BBa_K1144008) showing their fluorescent reporter, mCherry, after induction with 10 uM dexamethasone! The filter used was TRITC''']]
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====References====
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Aoyama, T. & Chua N. (1997). A glucocorticoid-mediated transcriptional induction system in transgenic plants. ''The Plant Journal, 11''(3): 605-612.
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Nickel Removal System
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==Nickel Removal System==
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Here are the parts sent to the registry:
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[[File:partsNi.jpg|700px|center|]]
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[[File:Prcna_hoxN.jpg|400px|thumb|center|Prcna-HoxN and HoxN-RFP]]
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Latest revision as of 03:40, 28 September 2013

Parts

Glucocorticoid sensor

Contents

Glucocorticoid sensor

Here are the parts sent to the registry:

PartsGC.jpg


Name table
Part Name Registry Number
E1 BBa_K1144001
E2 BBa_K1144002
E3 BBa_K1144003
E4 BBa_K1144004
E1GF BBa_K1144005
E2GF BBa_K1144006
E3GF BBa_K1144007
E4GF BBa_K1144008

You can see the confirmation gels of the parts below:


PCR confirmation: E1GF-E1GF(2)-E2GF-E2GF(2)-WM-E3GF-E3GF(2)-E4GF-E4GF(2)
Co-transformation: WM-E1GF2-E2GF-E3GF1-E3GF2-E4GF-WM- E3GF3-E4GF2-E2GF2
PCR confirmation of the parts cloned in the pSB1C3 backbone using the specifically designed primers 15 and 31 (see primers)


Characterization of reporters

To assess the strength of our promoters when induced with Dexamethasone, we performed a fluorometric assay using mCherry as our reporter.Since we don't have our transactivating protein ready yet, we co-transformed our parts into the E. coli DH10B strain with the pAT7002 vector (Aoyama and Chua, 1997), which contains a well characterized Glucocorticoid Responsive Element that also uses the GAL4 DNA binding domain.

Unsaturated curve where we see how the mCherry reporter appears after the addition of 10 uM dexamethasone, a glucocorticoid. Emmision intensity was measured at 607nm after excitation at 586nm


Saturated curve for fluorescence. After three hours of the addition of 10 uM dexamethasone the reporter (mCherry) reaches its highest signal. We can see the different strengths depending on the number of UAS boxes. Emmision intensity was measured at 607nm after excitation at 586nm


We also decided to visually inspect our induced transformants. Here two of the images taken using a epifluorescent microscopy with a TRITC filter.


Cells with the E2GF part (BBa_K1144006) showing their fluorescent reporter, mCherry, after induction with 10 uM dexamethasone! The filter used was TRITC
Cells with the E4GF part (BBa_K1144008) showing their fluorescent reporter, mCherry, after induction with 10 uM dexamethasone! The filter used was TRITC


References

Aoyama, T. & Chua N. (1997). A glucocorticoid-mediated transcriptional induction system in transgenic plants. The Plant Journal, 11(3): 605-612.

Nickel Removal System

Nickel Removal System

Here are the parts sent to the registry:

PartsNi.jpg
Prcna-HoxN and HoxN-RFP


Retrieved from "http://2013.igem.org/Team:Colombia_Uniandes/Parts"