Difference between revisions of "Part:BBa K3930028"
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                 <b>Figure 1: </b> <b> Integration of pCONCOMBRE in the cyanobacterium genome</b>
 
                 <b>Figure 1: </b> <b> Integration of pCONCOMBRE in the cyanobacterium genome</b>
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                 <p>pCONCOMBRE integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. 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)</p>
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                 <p>pCONCOMBRE integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right and the NEB 1 kb DNA ladder on the left (note that a different ladder is presented on the theoretical gel)</p>
 
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Revision as of 09:55, 15 October 2021


Integrative left site in the NSI locus of the S. elongatus genome Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 590
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 777
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 590
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 590
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

Integrative locus NSI left arm of Synechococcus elongatus genome comes from the plasmid pAM4951 (Taton et al. 2014). This part is flanking the insert in 5', and must be used with the integrative locus (BBa_K3930027) NSI right arm part in 3'.

Results

Integration of part (BBa_K3930026) into the cyanobacterium genome

The part (BBa_K3930026) was transformed into S.elongatus UTEX 2973 following the triparental conjugation protocol of Gale et al. (2019). The insert is flanked by parts (BBa_K3930027) and (BBa_K3930028). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype.



Figure 1: Integration of pCONCOMBRE in the cyanobacterium genome

pCONCOMBRE integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right and the NEB 1 kb DNA ladder on the left (note that a different ladder is presented on the theoretical gel)


The Integrative locus NSI left arm (BBa_K3930028) coupled with the integrative locus (BBa_K3930027) NSI right arm part are functional under those experimental conditions, even if the right size was not achieved.


References

  1. Gale GAR, Osorio AAS, Puzorjov A, Wang B, McCormick AJ. 2019. Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit. JoVE (Journal of Visualized Experiments).(152):e60451. doi:10.3791/60451.
  2. Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepes J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, et al. 2014. Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria. Nucleic Acids Res. 42(17):e136. doi:10.1093/nar/gku673.
    3.