Difference between revisions of "Part:BBa K3930022"
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                 <b>Figure 1: </b> <b> Integration of pVIOLETTE in the LycoYeast genome</b>
 
                 <b>Figure 1: </b> <b> Integration of pVIOLETTE in the LycoYeast genome</b>

Revision as of 22:11, 14 October 2021


Down 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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

Integrative locus down XII-4 of S. cerevisiae genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016). This part needs to flank the insertion in 3', and must be used with the integrative locus (BBa_K3930021) XII-4 up part in 5'.

Results

Integration of the part (BBa_K3930003) into the yeast genome

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 the part (BBa_K3930021) and (BBa_K3930022). Figure 1 shows the electrophoresis gel of PCR on colony to verify clones.



Figure 1: Integration of pVIOLETTE in the LycoYeast genome

pVIOLETTE was checked with agarose electrophoresis gel and revealed with EtBr. A theoretical gel is presented on the right of each gel and the NEB 1 kb DNA ladder on the left (note that a different ladder is presented on the theoretical gel)


The Integrative locus XII-4 down (BBa_K3930022) coupled with the integrative locus (BBa_K3930021) XII-4 up part work 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.