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Revision as of 10:17, 27 September 2024
ISU1
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Origin
Pichia pastoris; synthesized
Properties
ISU1 is a gene in the brewing yeast GS115. We have identified this gene for knockout purposes.
Experimental Approach (pSCm-ΔISU1)
We generated pSCm-ΔISU1 through the ligation of the pSCm backbone with the target gene ΔISU1, followed by transformation into competent E. coli DH5α. Fig. 2a illustrates the presence of individual colonies on the plate.
A colony PCR was conducted to validate the colonies. The results are depicted in Fig. 2b, which exhibits bands of 288 bp, aligning with the expected length, verifying the successful construction of the plasmid.
Furthermore, sequencing was performed on the colonies. As demonstrated in Fig. 2c, the ΔISU1 gene was seamlessly ligated to the backbone without any detectable mutations, confirming the accurate construction of the pSCm-ΔISU1 plasmid.
(a: pSCm-ΔISU1 colonies; b: the gel electrophoresis validation of colony PCR of pSCm-ΔISU1 colonies; c: sequencing result of pSCm-ΔISU1)