Difference between revisions of "Part:BBa K2142004"

 
(Information contributed by City of London UK (2021))
 
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<partinfo>BBa_K2142004 short</partinfo>
 
<partinfo>BBa_K2142004 short</partinfo>
  
This coding sequence was used to generate a higher mutation rate. Norma this protein is used as repairing system. With overexpression  its possible to reach a mutation rate which is 50x more than normal. Mutation will occur on plasmid and genom.
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The sequence encodes for the dam methylase. This metylase is normally important for methylating DNA after replication. In our project we over expressed the translation of the protein which leads to an mutation increase by the factor of 50. The part is no standard BioBrick and can be found in BBa_K2142005
  
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===Information contributed by City of London UK (2021)===
===Usage and Biology===
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Part information is collated here to help future users of the BioBrick registry.
  
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Metadata:
<span class='h3bb'>Sequence and Features</span>
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*'''Group:''' City of London UK 2021
<partinfo>BBa_K2142004 SequenceAndFeatures</partinfo>
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*'''Author:''' Lucas Ng
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*'''Summary:''' Added information collated from existing scientific studies
  
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dam causes DNA methylation within the sequence GATC, preventing cleavage by the MboI restriction endonuclease
===Functional Parameters===
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<ref>Geier, G. E., and P. Modrich. 1979. “Recognition Sequence of the Dam Methylase of Escherichia Coli K12 and Mode of Cleavage of Dpn I Endonuclease.” The Journal of Biological Chemistry 254 (4): 1408–13. https://pubmed.ncbi.nlm.nih.gov/368070.</ref>.
<partinfo>BBa_K2142004 parameters</partinfo>
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It is likely to act in postreplication mismatch repair
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<ref>Lu, A. L., S. Clark, and P. Modrich. 1983. “Methyl-Directed Repair of DNA Base-Pair Mismatches in Vitro.” Proceedings of the National Academy of Sciences 80 (15): 4639–43. https://doi.org/10.1073/pnas.80.15.4639.</ref>
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rather than as a member of a restriction modification system, despite sharing sequence specificity with several type II restriction methylases and endonucleases as part of the N(4)/N(6)-methyltransferase family
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<ref>Horton, John R., Xing Zhang, Robert M. Blumenthal, and Xiaodong Cheng. 2015. “Structures of Escherichia Coli DNA Adenine Methyltransferase (Dam) in Complex with a Non-GATC Sequence: Potential Implications for Methylation-Independent Transcriptional Repression.” Nucleic Acids Research 43 (8): 4296–4308. https://doi.org/10.1093/nar/gkv251.</ref>.
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Furthermore, it may also be involved in DNA replication
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<ref>Smith, D. W., A. M. Garland, G. Herman, R. E. Enns, T. A. Baker, and J. W. Zyskind. 1985. “Importance of State of Methylation of OriC GATC Sites in Initiation of DNA Replication in Escherichia Coli.” The EMBO Journal 4 (5): 1319–26. https://pubmed.ncbi.nlm.nih.gov/3891329.</ref>
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====Reaction====
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a 2'-deoxyadenosine in DNA + S-adenosyl-L-methionine = an N6-methyl-2'-deoxyadenosine in DNA + H(+) + S-adenosyl-L-homocysteine
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<ref>“Rhea - Annotated Reactions Database.” n.d. Rhea. Accessed July 26, 2021. https://www.rhea-db.org/rhea/15197.</ref>
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===References===

Latest revision as of 17:37, 26 July 2021


Dam methylase

The sequence encodes for the dam methylase. This metylase is normally important for methylating DNA after replication. In our project we over expressed the translation of the protein which leads to an mutation increase by the factor of 50. The part is no standard BioBrick and can be found in BBa_K2142005

Information contributed by City of London UK (2021)

Part information is collated here to help future users of the BioBrick registry.

Metadata:

  • Group: City of London UK 2021
  • Author: Lucas Ng
  • Summary: Added information collated from existing scientific studies

dam causes DNA methylation within the sequence GATC, preventing cleavage by the MboI restriction endonuclease [1].

It is likely to act in postreplication mismatch repair [2] rather than as a member of a restriction modification system, despite sharing sequence specificity with several type II restriction methylases and endonucleases as part of the N(4)/N(6)-methyltransferase family [3].

Furthermore, it may also be involved in DNA replication [4]

Reaction

a 2'-deoxyadenosine in DNA + S-adenosyl-L-methionine = an N6-methyl-2'-deoxyadenosine in DNA + H(+) + S-adenosyl-L-homocysteine [5]

References

  1. Geier, G. E., and P. Modrich. 1979. “Recognition Sequence of the Dam Methylase of Escherichia Coli K12 and Mode of Cleavage of Dpn I Endonuclease.” The Journal of Biological Chemistry 254 (4): 1408–13. https://pubmed.ncbi.nlm.nih.gov/368070.
  2. Lu, A. L., S. Clark, and P. Modrich. 1983. “Methyl-Directed Repair of DNA Base-Pair Mismatches in Vitro.” Proceedings of the National Academy of Sciences 80 (15): 4639–43. https://doi.org/10.1073/pnas.80.15.4639.
  3. Horton, John R., Xing Zhang, Robert M. Blumenthal, and Xiaodong Cheng. 2015. “Structures of Escherichia Coli DNA Adenine Methyltransferase (Dam) in Complex with a Non-GATC Sequence: Potential Implications for Methylation-Independent Transcriptional Repression.” Nucleic Acids Research 43 (8): 4296–4308. https://doi.org/10.1093/nar/gkv251.
  4. Smith, D. W., A. M. Garland, G. Herman, R. E. Enns, T. A. Baker, and J. W. Zyskind. 1985. “Importance of State of Methylation of OriC GATC Sites in Initiation of DNA Replication in Escherichia Coli.” The EMBO Journal 4 (5): 1319–26. https://pubmed.ncbi.nlm.nih.gov/3891329.
  5. “Rhea - Annotated Reactions Database.” n.d. Rhea. Accessed July 26, 2021. https://www.rhea-db.org/rhea/15197.