Difference between revisions of "Part:BBa K3930021"
Line 39: Line 39:
 
</ol>
 
</ol>
 
</html>
 
</html>
−
Integrative locus XII-4 up of S. cerevisiae genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016). This part needs to flank the insertion in 5', and must be used with the integrative locus (BBa_K39300022) XII-4 down part in 3'.
+
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 13:57, 11 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

Integrative locus up 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 5', and must be used with the integrative locus (BBa_K3930022) XII-4 up part in 3'.

Results

Integration of the part (BBa_K3930001) 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_K39300021) and (BBa_K39300022). 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 up (BBa_K3930021) coupled with the integrative locus (BBa_K3930022) XII-4 down 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.