Difference between revisions of "Part:BBa K5034227:Design"

 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
aaa
 
−
 
  
 +
PPK2 gene was PCR-amplified and inserted into the plasmid pYYDT(after NdeI and XhoI digestion) to get the expression vector. It then got transformed into E. coli DH5α to amplify, and was purified by picking monoclonal culture and sequencing.
  
 
===Source===
 
===Source===
  
−
aaa
+
Polyphosphate kinase 2(PPK2) from <i>Pseudomonas paraeruginosa</i>. NCBI reference sequence: NZ_CP020560.1:c164262-163366
  
 
===References===
 
===References===
 +
 +
Zhang, H., Ishige, K., & Kornberg, A. (2002). A polyphosphate kinase (PPK2) widely conserved in bacteria. Proceedings of the National Academy of Sciences, 99(26), 16678-16683. doi:10.1073/pnas.262655199
 +
 +
Neville, N., Roberge, N., & Jia, Z. (2022). Polyphosphate Kinase 2 (PPK2) Enzymes: Structure, Function, and Roles in Bacterial Physiology and Virulence. International Journal of Molecular Sciences, 23(2), 670. doi:10.3390/ijms23020670

Revision as of 07:34, 29 September 2024


Pi <-> PolyP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 11
    Illegal PstI site found at 3787
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 11
    Illegal PstI site found at 3787
    Illegal NotI site found at 2828
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 11
    Illegal BglII site found at 3574
    Illegal XhoI site found at 4985
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 11
    Illegal PstI site found at 3787
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 11
    Illegal PstI site found at 3787
    Illegal NgoMIV site found at 556
    Illegal NgoMIV site found at 4238
    Illegal NgoMIV site found at 4521
    Illegal AgeI site found at 396
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3150
    Illegal SapI.rc site found at 4087
    Illegal SapI.rc site found at 4297


Design Notes

PPK2 gene was PCR-amplified and inserted into the plasmid pYYDT(after NdeI and XhoI digestion) to get the expression vector. It then got transformed into E. coli DH5α to amplify, and was purified by picking monoclonal culture and sequencing.

Source

Polyphosphate kinase 2(PPK2) from Pseudomonas paraeruginosa. NCBI reference sequence: NZ_CP020560.1:c164262-163366

References

Zhang, H., Ishige, K., & Kornberg, A. (2002). A polyphosphate kinase (PPK2) widely conserved in bacteria. Proceedings of the National Academy of Sciences, 99(26), 16678-16683. doi:10.1073/pnas.262655199

Neville, N., Roberge, N., & Jia, Z. (2022). Polyphosphate Kinase 2 (PPK2) Enzymes: Structure, Function, and Roles in Bacterial Physiology and Virulence. International Journal of Molecular Sciences, 23(2), 670. doi:10.3390/ijms23020670