Regulatory

Part:BBa_K2703002:Experience

Designed by: Aurelie Bouin, Victor Sayous   Group: iGEM18_Sorbonne_U_Paris   (2018-10-01)
Revision as of 15:00, 9 October 2018 by Saniya kari (Talk | contribs)


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Applications of BBa_K2703002

Our part is compatible with the PhytoBrick MoClo standard. This PhytoBrick was intented to be use as constitutive promoter for the transcription unit of paromomycine in the CARGO of our synthetic retrotransposons for directed evolution on the Chlamydomoas. It is part of our reporter system to characterize the activity and transposition rate.

We couldn’t directly use our Phytobrick for the characterisation because we didn’t have time to construct all the other that are needed for the final assembly. Instead we use a slightly different Phytobrick with the same P PSAD. sequence but with one different fusion site. This part was assembled in a functional Transcription Unit with the resistance gene paromomycine that was as a reporter gene. After transformation in C.reinhardtii D66 the functionality of the promoter was tested by counting the number of chlamydomonas colony resistant to paromomycine (15 ug/ml). Different constructions were made to characterize and compare the functionality of P pSAD . We tested different combination of promoters and 3’UTR.

Name F1 Prom F2 5'UTR F3 Resistance F4 3'UTR F5
pCM-1 GGAG PAR TACT 5’UTRAR AATG Paromycin GCTT TRBCS2 CGCT
pCM-2 GGAG PAR TACT 5’UTRAR AATG Paromycin GCTT TPSAD CGCT
pCM-3 GGAG PPSAD TACT 5’UTRPSAD AATG Paromycin GCTT TRBCS2 CGCT
pCM-4 GGAG PPSAD TACT 5’UTRPSAD AATG Paromycin GCTT TPSAD CGCT
Figure 2: The 4 different devices made to test the functionality of the promoter PPSAD. The construction were made with the MoClo kit made for C.reinhardtii by P.Crozet et al 2018 1.

User Reviews

UNIQ3c1d9917e0e0e019-partinfo-00000002-QINU UNIQ3c1d9917e0e0e019-partinfo-00000003-QINU