Difference between revisions of "Part:BBa K3930027"
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<h3>Integration of part (BBa_K3930026) into the cyanobacterium genome</h3>
 
<h3>Integration of part (BBa_K3930026) into the cyanobacterium genome</h3>
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<p> The part (BBa_K3930026) was transformed into the <i>S.elongatus</i> UTEX 2973 following the triparental conjugation  protocol of Gale et al. (2019). The construction is flanked by the part (BBa_K3930027) and (BBa_K3930028). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype.</p>
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<p> The part (BBa_K3930026) was transformed into the <i>S.elongatus</i> UTEX 2973 following the triparental conjugation  protocol of Gale et al. (2019). The construction is flanked by parts (BBa_K3930027) and (BBa_K3930028). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype.</p>
 
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                 <b>Figure 1: </b> <b> Integration of pCONCOMBRE in the cyanobacterium genome</b>
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                 <b>Figure 1: </b> <b> Integration of pCONCOMBRE insert 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 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 insert 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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<p><b>The integrative locus NSI right arm (BBa_K3930027) coupled with the integrative locus (BBa_K3930028) NSI left arm part are functional under those experimental conditions.</b><p>
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<p><b>The integrative locus NSI right arm (BBa_K3930027) coupled with the integrative locus (BBa_K3930028) NSI left arm part are functional under these experimental conditions.</b><p>
 
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<h2>References</h2>
 
<h2>References</h2>

Revision as of 13:10, 15 October 2021


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 441
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 441
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 441
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 441
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 651

Introduction

Integrative locus NSI right arm of Synechococcus elongatus genome comes from the plasmid pAM4951 (Taton et al. 2014). This part is flanking the insert in 3', and must be used with the integrative locus (BBa_K3930028) NSI left arm part in 5'.

Results

Integration of part (BBa_K3930026) into the cyanobacterium genome

The part (BBa_K3930026) was transformed into the S.elongatus UTEX 2973 following the triparental conjugation protocol of Gale et al. (2019). The construction 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 insert in the cyanobacterium genome

pCONCOMBRE insert 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 right arm (BBa_K3930027) coupled with the integrative locus (BBa_K3930028) NSI left arm part are functional under these experimental conditions.


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.