Difference between revisions of "Part:BBa K5049007"
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| + | Pir1 is another anchor protein of Saccharomyces cerevisiae, widely utilized in yeast display systems. To explore its potential compared to GCW61, we aimed to construct a Xylanase-Pir1 fusion protein. Using the same gene expression strategy as with GCW61, the Pir1 sequence from Saccharomyces cerevisiae was synthesized with a GS linker at the N-terminus, flanked by EcoRI-XbaI-NgoMIV as a prefix and AgeI-SpeI-PstI as a suffix by IDT. This basic part, GS-Pir1, was assembled into the pSB1C3 vector (BBa_AAAA), and the composite part, PGTH1-Xylanase-Pir1/pSB1C3, was then created (BBa_BBBB). To evaluate its performance in yeast and compare it with the GCW61 anchor, the PGTH1-Xylanase-Pir1 construct was cloned into the pZAHR vector. The construct was verified by colony PCR (Figure 3) and confirmed by DNA sequencing. | ||
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Revision as of 03:12, 10 September 2024
PGTH1-Xylanase-Pir1
CONSTRUCTION
Pir1 is another anchor protein of Saccharomyces cerevisiae, widely utilized in yeast display systems. To explore its potential compared to GCW61, we aimed to construct a Xylanase-Pir1 fusion protein. Using the same gene expression strategy as with GCW61, the Pir1 sequence from Saccharomyces cerevisiae was synthesized with a GS linker at the N-terminus, flanked by EcoRI-XbaI-NgoMIV as a prefix and AgeI-SpeI-PstI as a suffix by IDT. This basic part, GS-Pir1, was assembled into the pSB1C3 vector (BBa_AAAA), and the composite part, PGTH1-Xylanase-Pir1/pSB1C3, was then created (BBa_BBBB). To evaluate its performance in yeast and compare it with the GCW61 anchor, the PGTH1-Xylanase-Pir1 construct was cloned into the pZAHR vector. The construct was verified by colony PCR (Figure 3) and confirmed by DNA sequencing.
===Sequence and Features===