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Part:BBa_I52002:Design

Designed by Reshma Shetty   Group: Knight Lab   (2008-01-05)

From partsregistry.org

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#Ioannou-Nat-Genet-1994 pmid=8136839
#Ioannou-Nat-Genet-1994 pmid=8136839
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*US Patent Number 5,910,438 "Cloning and/or sequencing vector" Bernard P, Gabant P, University Libre de Bruxelles, 1999. [http://www.google.com/patents?vid=USPAT5910438 Google Patents]
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*US Patent Number 6,180,407 B1 "Cloning and/or sequencing vector" Bernard P, Gabant P, Universit Libre de Bruxelles, 2001. [http://www.google.com/patents?vid=USPAT6180407 Google Patents]
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*US Patent Number 7,176,029 B2 "Cloning and/or sequencing vector" Bernard P, Gabant P, Universit Libre de Bruxelles, 2007. [http://www.google.com/patents?vid=USPAT7176029 Google Patents]

Revision as of 15:52, 10 March 2010

ccdB and minimal pUC19 derived high copy origin

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 435
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]


Design Notes

The BioBrick standard vectors are designed to be easy to use for their most common purpose: assembly of BioBrick standard biological parts. To meet this requirement, we included BBa_P1016 within the BioBrick cloning site. BBa_P1016 encodes the positive selection marker ccdB. Positive selection markers prevent one of the most common problems during assembly of BioBrick parts: contamination of the ligation reaction with uncut plasmid DNA[1]. Any cells transformed with the uncut plasmid DNA produce the lethal protein ccdB and die[2, 3, 4]. Note, however, that the drawback of this solution is that inclusion of this part requires that users propagate both the base vector and any derived vectors in E. coli strains tolerant of ccdB expression such as DB3.1[5, 6].

BioBrick standard vectors are also designed to be easy to purify. To meet this requirement, we included a pUC19-derived origin (BBa_I50022) in addition to the ccdB selection marker within the BioBrick cloning site of BioBrick standard vectors[7, 8, 9]. The high copy origin encoded by BBa_I50022 means that both base vector DNA and any derived vector DNA are easily purified in large quantities, irrespective of whether the vector replication origin is low copy or not[10, 11]. Cloning a BioBrick part into the BioBrick cloning site removes the high copy origin in the cloning site thereby restoring replication control to the vector origin.

Source

BBa_P1016 (ccdB positive selection marker) and BBa_I50022 (minimal high copy origin derived from pUC19).

References

  1. Bernard P, Gabant P, Bahassi EM, and Couturier M. Positive-selection vectors using the F plasmid ccdB killer gene. Gene 1994 Oct 11; 148(1) 71-4. pmid:7926841. PubMed HubMed [Bernard-Gene-1994]
  2. Bernard P. New ccdB positive-selection cloning vectors with kanamycin or chloramphenicol selectable markers. Gene 1995 Aug 30; 162(1) 159-60. pmid:7557407. PubMed HubMed [Bernard-Gene-1995]
  3. Bernard P. Positive selection of recombinant DNA by CcdB. Biotechniques 1996 Aug; 21(2) 320-3. pmid:8862819. PubMed HubMed [Bernard-Biotechniques-1996]
  4. Bernard P and Couturier M. Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes. J Mol Biol 1992 Aug 5; 226(3) 735-45. pmid:1324324. PubMed HubMed [Bernard-J-Mol-Biol-1992]
  5. Miki T, Park JA, Nagao K, Murayama N, and Horiuchi T. Control of segregation of chromosomal DNA by sex factor F in Escherichia coli. Mutants of DNA gyrase subunit A suppress letD (ccdB) product growth inhibition. J Mol Biol 1992 May 5; 225(1) 39-52. pmid:1316444. PubMed HubMed [Miki-J-Mol-Biol-1992]
  6. Vieira J and Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 1982 Oct; 19(3) 259-68. pmid:6295879. PubMed HubMed [Vieira-Gene-1982]
  7. Norrander J, Kempe T, and Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 1983 Dec; 26(1) 101-6. pmid:6323249. PubMed HubMed [Norrander-Gene-1983]
  8. Yanisch-Perron C, Vieira J, and Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 1985; 33(1) 103-19. pmid:2985470. PubMed HubMed [Yanisch-Perron-Gene-1985]
  9. Cabello F, Timmis K, and Cohen SN. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature 1976 Jan 29; 259(5541) 285-90. pmid:765836. PubMed HubMed [Cabello-Nature-1976]
  10. Ioannou PA, Amemiya CT, Garnes J, Kroisel PM, Shizuya H, Chen C, Batzer MA, and de Jong PJ. A new bacteriophage P1-derived vector for the propagation of large human DNA fragments. Nat Genet 1994 Jan; 6(1) 84-9. doi:10.1038/ng0194-84 pmid:8136839. PubMed HubMed [Ioannou-Nat-Genet-1994]
All Medline abstracts: PubMed HubMed
  • US Patent Number 5,910,438 "Cloning and/or sequencing vector" Bernard P, Gabant P, University Libre de Bruxelles, 1999. Google Patents
  • US Patent Number 6,180,407 B1 "Cloning and/or sequencing vector" Bernard P, Gabant P, Universit Libre de Bruxelles, 2001. Google Patents
  • US Patent Number 7,176,029 B2 "Cloning and/or sequencing vector" Bernard P, Gabant P, Universit Libre de Bruxelles, 2007. Google Patents
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