Part:BBa_K5508007
Gal1,10-mCherry-CYC1
The fluorescent protein gene BBa_K4335004 (mCherry) was placed under the control of the BBa_K5477005 promoter (Gal1, 10), and its transcription was terminated by BBa_K3384311(CYC1) to validate the availability of theSaccharomyces cerevisiae's protein expression system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 374
- 1000COMPATIBLE WITH RFC[1000]
Results
(1)Plasmid Construction
The construction process used the following primers mCherry-Gibson-F (CACTATAGGGCCCGGGCGTCGACatggtgagcaagggcgaggag), and mCherry-Gibson-R (GCTAGCCGCGGTACCAAGCTTttacttgtacagctcgtcc) to amplify the mCherry gene fragments, which were later assembled using Gibson. SalI and HindIII were used to linearise the carrier. The vectors were linearised with SalI and HindIII, followed by Gibson assembly to ligate the vectors to the target genes.
(2)Fluorescence detection
After the correctness of the recombinant plasmid was verified, it was transferred into BY4741 for culture. After 48 hours of adding the inducer galactose, the results are shown. The bacterial fluid appeared a light pink color, proving that our expression system was successful.
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