Difference between revisions of "Part:BBa K5351003"
 
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         <img src="https://static.igem.wiki/teams/5351/bba-k5351003/1.png" width="70%" alt="Figure 1. Gene map of ISU1">
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             <caption>Figure 1. Gene map of ISU1</caption>
 
             <caption>Figure 1. Gene map of ISU1</caption>
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         <img src="https://static.igem.wiki/teams/5351/bba-k5351003/2.png" width="70%" alt="Figure 2. The Monoclonal antibody validation and sequencing of pSCm-ΔISU1 (DH5α)">
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         <img src="https://static.igem.wiki/teams/5351/bba-k5351003/2.png" width="50%" alt="Figure 2. The Monoclonal antibody validation and sequencing of pSCm-ΔISU1 (DH5α)">
 
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             <caption>Figure 2. The Monoclonal antibody validation and sequencing of pSCm-ΔISU1 (DH5α)  
 
             <caption>Figure 2. The Monoclonal antibody validation and sequencing of pSCm-ΔISU1 (DH5α)  

Latest revision as of 06:30, 29 September 2024

ISU1


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

BBa_K5351001 Documentation

Origin

Pichia pastoris; synthesized

Properties

ISU1 is a gene in the brewing yeast GS115. We have identified this gene for knockout purposes.

Figure 1. Gene map of ISU1
Figure 1. Gene map of ISU1

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.

Figure 2. The Monoclonal antibody validation and sequencing of pSCm-ΔISU1 (DH5α)
Figure 2. The Monoclonal antibody validation and sequencing of pSCm-ΔISU1 (DH5α)
(a: pSCm-ΔISU1 colonies; b: the gel electrophoresis validation of colony PCR of pSCm-ΔISU1 colonies; c: sequencing result of pSCm-ΔISU1)