Difference between revisions of "Part:BBa K2703002:Experience"

Revision as of 15:00, 9 October 2018

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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

UNIQ67b98d0c37f08c30-partinfo-00000002-QINU UNIQ67b98d0c37f08c30-partinfo-00000003-QINU

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