Escherichia coli/Vectors

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(Stringent vs. relaxed replication)
 
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===Stringent vs. relaxed replication===
===Stringent vs. relaxed replication===
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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 abscence 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.--[[User:Bcanton|BC]] 19:05, 3 Sep 2005 (EDT)
+
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 absence 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.--[[User:Bcanton|BC]] 19:05, 3 Sep 2005 (EDT)
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===Nomenclature===
 +
<biblio>
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#Novick-BacteriolRev-1976 pmid=1267736
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</biblio>
==Common vectors==
==Common vectors==
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|-
|-
|pACYC and its derivatives
|pACYC and its derivatives
-
|p15A<cite>Chang-JBacteriol-1978</cite>
+
|[[p15A]]<cite>Chang-JBacteriol-1978</cite>
|10-12
|10-12
|-
|-
-
|pSC101 and its derivatives
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|[[pSC101]] and its derivatives
|pSC101<cite>Cohen-JBacteriol-1977</cite>
|pSC101<cite>Cohen-JBacteriol-1977</cite>
|5
|5
|-
|-
-
|ColE1
+
|[[ColE1]]
|ColE1<cite>Bazaral-J-Mol-Biol-1968,Hershfield-Proc-Natl-Acad-Sci-USA-1974</cite>
|ColE1<cite>Bazaral-J-Mol-Biol-1968,Hershfield-Proc-Natl-Acad-Sci-USA-1974</cite>
|15-20
|15-20
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*pSCANS [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Nucleotide&cmd=Retrieve&list_uids=6689911&dopt=GenBank genbank] [http://genome.bnl.gov/Vectors/pscans.php vector info] [http://www.genome.bnl.gov/Protocols/pscans.shtml cookbook]
*pSCANS [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Nucleotide&cmd=Retrieve&list_uids=6689911&dopt=GenBank genbank] [http://genome.bnl.gov/Vectors/pscans.php vector info] [http://www.genome.bnl.gov/Protocols/pscans.shtml cookbook]
*[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=42704 pSC101 replication origin]
*[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=42704 pSC101 replication origin]
 +
*[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=208958 pBR322]
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*[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=58255 pACYC184]
==References==
==References==
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// Discusses transferable drug resistance in ''Escherichia coli'' (i.e. plasmids with resistance markers)
// Discusses transferable drug resistance in ''Escherichia coli'' (i.e. plasmids with resistance markers)
#Cohen-Proc-Natl-Acad-Sci-USA-1973a pmid=4576014
#Cohen-Proc-Natl-Acad-Sci-USA-1973a pmid=4576014
-
// Discusses generation of plasmids by sharing DNA, their transformation and replication.
+
// Discusses generation of plasmids by shearing DNA, their transformation and replication.
#Cohen-Proc-Natl-Acad-Sci-USA-1973b pmid=4594039
#Cohen-Proc-Natl-Acad-Sci-USA-1973b pmid=4594039
// Discusses use of vector to propagate other DNA fragments
// Discusses use of vector to propagate other DNA fragments
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**table with links to properties/sequences of pET vectors
**table with links to properties/sequences of pET vectors
**note: slow to load
**note: slow to load
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 +
*[[Ecoliwiki:Category:Plasmids|Plasmids at EcoliWiki.org]]
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**EcoliWiki.org has entered many plasmids that are available for annotation. Included are plasmid maps, features and sequences.
==See also==
==See also==
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*[[Vectors]]
*[[Vectors]]
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[[Category:Escherichia coli]] [[Category:DNA]]
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[[Category:Escherichia coli]] [[Category:DNA]] [[Category:Vectors]]

Current revision

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 absence 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

  1. Novick RP, Clowes RC, Cohen SN, Curtiss R 3rd, Datta N, and Falkow S. . pmid:1267736. PubMed HubMed [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

  1. Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, and Falkow S. . pmid:344137. PubMed HubMed [Bolivar-Gene-1977]
    vector pBR322

  2. Chang AC and Cohen SN. . pmid:149110. PubMed HubMed [Chang-JBacteriol-1978]
    p15A origin

  3. Cohen SN and Chang AC. . pmid:334752. PubMed HubMed [Cohen-JBacteriol-1977]
    pSC101 origin

  4. Bazaral M and Helinski DR. . pmid:4939624. PubMed HubMed [Bazaral-J-Mol-Biol-1968]
    original discovery of ColE1 plasmid

  5. Hershfield V, Boyer HW, Yanofsky C, Lovett MA, and Helinski DR. . pmid:4610576. PubMed HubMed [Hershfield-Proc-Natl-Acad-Sci-USA-1974]
    use of ColEI as a vector

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

  8. Cohen SN and Chang AC. . pmid:4576014. PubMed HubMed [Cohen-Proc-Natl-Acad-Sci-USA-1973a]
    Discusses generation of plasmids by shearing DNA, their transformation and replication.

  9. Cohen SN, Chang AC, Boyer HW, and Helling RB. . pmid:4594039. PubMed HubMed [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

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