Difference between revisions of "Part:BBa K1362401:Experience"

 
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===Applications of BBa_K1362401===
 
===Applications of BBa_K1362401===
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===Characterization in <i>Nicotiana benthamiana</i>===
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<b>Group</b>: [http://2016.igem.org/Team:Valencia_UPV Valencia_UPV team 2016]
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<b>Author</b>: Valencia UPV iGEM team (Iván Casas Rodrigo, Mónica Victoria Gutiérrez Salazar, Alicia Climent Catalá, Álvaro Ballesteros González)
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<b>Summary</b>
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Valencia UPV 2016 team tested and demonstrated the functionality of inteins in a plant chassis, specifically in <i>Nicotiana benthamiana</i>. We used the inteins with our split-Cas9 ([https://parts.igem.org/Part:BBa_K2017000 BBa_K2017000] and [https://parts.igem.org/Part:BBa_K2017001 BBa_K2017001]). The inteins allowed to fusion of the two parts of the split-Cas9 in vivo, in order to obtain a fully functional Cas9 protein. In Figure 1 it is shown an electrophoresis gel with the results of the testing of split-Cas9 in plants. For further information about the experimental design, check the information of split-Cas9 parts, [https://parts.igem.org/Part:BBa_K2017000 BBa_K2017000] and [https://parts.igem.org/Part:BBa_K2017001 BBa_K2017001]
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[[Image:BBa_K201700_digestionsplitcas9.jpg|600px|thumb|center|'''Figure 1:''' Electrophoresis gel of XT1 amplicons digestion. Mutation efficiencies of Split-intein-Cas9 system and classical Cas9 system are compared by resistant bands signal intensity.]]
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===Additional information===
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*Team:iGEM-IISERPUNE-2020
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*We aimed to use this part for circularization of the cyclotide protein. To build the biobrick we preferred restriction-free cloning but we realised that the biobrick was not compatible with restriction-free cloning. So we modified the part and built the modified version of C- Intein which is compatible with restriction-free cloning technique: BBa_K3582020.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 18:14, 24 October 2020


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_K1362401


Characterization in Nicotiana benthamiana

Group: [http://2016.igem.org/Team:Valencia_UPV Valencia_UPV team 2016]

Author: Valencia UPV iGEM team (Iván Casas Rodrigo, Mónica Victoria Gutiérrez Salazar, Alicia Climent Catalá, Álvaro Ballesteros González)

Summary

Valencia UPV 2016 team tested and demonstrated the functionality of inteins in a plant chassis, specifically in Nicotiana benthamiana. We used the inteins with our split-Cas9 (BBa_K2017000 and BBa_K2017001). The inteins allowed to fusion of the two parts of the split-Cas9 in vivo, in order to obtain a fully functional Cas9 protein. In Figure 1 it is shown an electrophoresis gel with the results of the testing of split-Cas9 in plants. For further information about the experimental design, check the information of split-Cas9 parts, BBa_K2017000 and BBa_K2017001

Figure 1: Electrophoresis gel of XT1 amplicons digestion. Mutation efficiencies of Split-intein-Cas9 system and classical Cas9 system are compared by resistant bands signal intensity.


Additional information

  • Team:iGEM-IISERPUNE-2020
  • We aimed to use this part for circularization of the cyclotide protein. To build the biobrick we preferred restriction-free cloning but we realised that the biobrick was not compatible with restriction-free cloning. So we modified the part and built the modified version of C- Intein which is compatible with restriction-free cloning technique: BBa_K3582020.

User Reviews

UNIQ3c254fa98bfb5c9b-partinfo-00000000-QINU UNIQ3c254fa98bfb5c9b-partinfo-00000001-QINU