Escherichia coli/Vectors

From OpenWetWare

Jump to: navigation, search

Contents

General information

Stringent vs. relaxed replication

Plasmid replication control is usually controlled by balancing the levels of a positive and a negative regulator of replication. For some plasmids (pMB1/colE1 replicons) the positive regulator is an RNA and in others (e.g. pSC101) it is a protein. Plasmids with a protein positive regulator will not replicate in the absence of protein production - stringent control (although not the same as the stringent response due to a shortage of loaded tRNAs). Plasmids with an RNA positive regulator will continue to replicate in the abscence of protein production. This is termed relaxed control. High yields of plasmid may be obtained by halting protein production (via chloroamphenicol) when the culture reaches a high density and then continuing incubation for a number of hours. This might be of practical relevance when prepping the 1 and 3 series of Synthetic Biology plasmids.--BC 19:05, 3 Sep 2005 (EDT)

Nomenclature

Error fetching PMID 1267736:
  1. Error fetching PMID 1267736: [Novick-BacteriolRev-1976]

Common vectors

Plasmid Replicon Copy number
pBR322[2] and its derivatives pMB1 15-20
pUC vectors pMB1 500-700
pACYC and its derivatives p15A[3] 10-12
pSC101 and its derivatives pSC101[4] 5
ColE1 ColE1[5, 6] 15-20

Replicon compatibility

Incompatibility group Negative control element Comment
colE1[5, 6], pMB1 RNAI controls processing of pre-RNAII into primer. pUC is derived from pBR322 (a single mutation in the pBR322 Primer RNA and deletion of the rop gene) which is derived from a pMB1 replicon, and cannot co-reside with the colE1 incompatibility group.
IncFII, pT181 RNA controls synthesis of RepA protein
P1, F, R6K, pSC101[4], p15A[3] iterons sequesters RepA protein

Replication origins in different incompatibility groups are compatible. Replication origins in the same incompatibility group are not.

Some sets of vectors with compatible origins are available as a part of the Novagen Duet system. (from TK)

Individual vector links

Note: searching for cloning vector <insert vector name> when looking for vector sequences in NCBI Entrez Nucleotide search. It helps to cut down on the number of hits.

References

Error fetching PMID 4879515:
Error fetching PMID 4576014:
Error fetching PMID 4594039:
Error fetching PMID 149110:
Error fetching PMID 334752:
Error fetching PMID 344137:
Error fetching PMID 4939624:
Error fetching PMID 4610576:
  1. Error fetching PMID 344137: [Bolivar-Gene-1977]
    vector pBR322

  2. Error fetching PMID 149110: [Chang-JBacteriol-1978]
    p15A origin

  3. Error fetching PMID 334752: [Cohen-JBacteriol-1977]
    pSC101 origin

  4. Error fetching PMID 4939624: [Bazaral-J-Mol-Biol-1968]
    original discovery of ColE1 plasmid

  5. Error fetching PMID 4610576: [Hershfield-Proc-Natl-Acad-Sci-USA-1974]
    use of ColEI as a vector

  6. isbn:0-87969-577-3. [MolecularCloning]
  7. Error fetching PMID 4879515: [Anderson-Annu-Rev-Microbiol-1968]
    Discusses transferable drug resistance in Escherichia coli (i.e. plasmids with resistance markers)

  8. Error fetching PMID 4576014: [Cohen-Proc-Natl-Acad-Sci-USA-1973a]
    Discusses generation of plasmids by shearing DNA, their transformation and replication.

  9. Error fetching PMID 4594039: [Cohen-Proc-Natl-Acad-Sci-USA-1973b]
    Discusses use of vector to propagate other DNA fragments

All Medline abstracts: PubMed HubMed

External links

  • Plasmids at EcoliWiki.org
    • EcoliWiki.org has entered many plasmids that are available for annotation. Included are plasmid maps, features and sequences.

See also

Personal tools