Difference between revisions of "Part:BBa K3930021"
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<h3>Integration of part (BBa_K3930001) into the yeast genome at locus XII-4</h3>
 
<h3>Integration of part (BBa_K3930001) into the yeast genome at locus XII-4</h3>
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<p> The part (BBa_K3930003) was linearized and transformed into the <i>S. cerevisiae</i> LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The construction is flanked by parts (BBa_K39300021) and (BBa_K39300022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype.</p>
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<p> The part (BBa_K3930003) was linearized and transformed into the <i>S. cerevisiae</i> LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The construction is flanked by parts (BBa_K39300021) and (BBa_K39300022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).</p>
 
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Revision as of 18:37, 15 October 2021


Up integrative sequence to target locus XII-4 of Saccharomyces cerevisiae genome Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 73
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

The sequence to target the locus XII-4 of S. cerevisiae genome come from the EasyClone-MarkerFree kit (Jessop-Fabre et al., 2016). This part is flanking the insert in 5' and must be used with the (BBa_K3930022) XII-4 up part in 3'.

Results

Integration of part (BBa_K3930001) into the yeast genome at locus XII-4

The part (BBa_K3930003) was linearized and transformed into the S. cerevisiae LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The construction is flanked by parts (BBa_K39300021) and (BBa_K39300022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).



Figure 1: Integration of pVIOLETTE insert in the LycoYeast genome at locus XII-4

pVIOLETTE insert integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right (note that a different ladder is presented on the theoretical gel)


The integrative locus XII-4 up (BBa_K3930021) coupled with the integrative locus (BBa_K3930022) XII-4 down part is functional to target integration at the XII-4 locus under those experimental conditions.


References

  1. Chen X, Shukal S, Zhang C. 2019. Integrating Enzyme and Metabolic Engineering Tools for Enhanced α-Ionone Production. J Agric Food Chem. 67(49):13451–13459. doi:10.1021/acs.jafc.9b00860.
    2.