DH5-Alpha E.coli: Difference between revisions

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A Microbial Biorealm page on the genus BI246 General Microbiology Skidmore College Fall 2011
[[Category:Short pages]]
Contents
{{Uncurated}}
 
[[Category:Pages edited by students of Prof. McDevitt‏‎]]
    1 Classification
    2 Description and significance
    3 Genome structure
    4 Cell structure, metabolism & life cycle
    5 Ecology (including pathogenesis)
    6 Interesting feature
    7 References
 
Classification
Classification


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Genus: Escherichia
Genus: Escherichia
Species:  Escherishia coli DH5-Alpha
Species:  Escherishia coli DH5-Alpha
(1)




Description and significance


Description and significance
This strain of E. Coli is not a pathogen, and was developed for laboratory cloning use.  This strain was developed by D. Hanahan as a cloning strain with multiple mutations that enable high-efficiency transformations (2).  The mutations that the DH5-Alpha strain has are:  dlacZ Delta M15 Delta(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 (2).  These mutations correspond to the distinct characteristics that make the DH5-Alpha strain excel in laboratory cloning procedures (2). 


This strain of E. Coli is not a pathogen, and was developed for laboratory cloning useThis strain was developed by D. Hanahan as a cloning strain with multiple mutations that enable high-efficiency transformations (Cite PDF).  The mutations that the DH5-Alpha strain has are:  dlacZ Delta M15 Delta(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 (cite PDF).  These mutations correspond to the distinct characteristics that make the DH5-Alpha strain excel in laboratory cloning procedures. 
lacZ Delta M15 mutation:  Allows for blue-white screening for recombinant cells.  (3)
endA1 mutation:  Allows for lower endonuclease degradation which ensures higher plasmid transfer rates.  (4)
recA1 mutation: reduces homologous recombination for a more stable insert.  (5)


lacZ Delta M15 mutation:  Allows for blue-white screening for recombinant cells.  (http://bloodjournal.hematologylibrary.org/content/71/2/463.short)
endA1 mutation:  Allows for lower endonuclease degradation which ensures higher plasmid transfer rates.  (http://www.ncbi.nlm.nih.gov/pubmed/18245285)
recA1 mutation: reduces homologous recombination for a more stable insert.  (http://www.ncbi.nlm.nih.gov/pubmed/6363880)




Genome structure


The genomic structure of this strain is a singular circular chromosome consisting of 4,686,137 nucleotides, 4359 genes, and 4128 protein encoding genes (6)


Describe the disease caused by this organism if it is a pathogen, or the natural macroscopic "field guide" appearance and habitat of your organism if it is not. What is or has been the impact your organism on human history or our environment?. How does it do this? How have we harnessed this power, or tried to prevent it? In other words, how do you know it if you see it, and how does its presence influence humans in the present, and historically?
This strain also contains plasmids, and has the ability to accept plasmid insertion exceptionally well.


Genome structure


The genomic structure of this strain is a singular circular chromosome consisting of 4,686,137 nucleotides, 4359 genes, and 4128 protein encoding genes (Cite http://www.ncbi.nlm.nih.gov/sites/entrez!db=genome&cmd=Retrieve&dopt=Overview&list_uids=22068)


Describe the size and content of the genome. How many chromosomes and plasmids? Circular or linear? Other interesting features? What is known about its sequence?
Cell structure, metabolism & life cycle
Cell structure, metabolism & life cycle


Provide a physical and biochemical description of the organism. What kind of organism is it, what does it look like, how is it built, what are its metabolic properties, how can it be identified, what is it's life cycle, &c. In other words, describe the organism from its perspective.
E. coli are gram-negative bacillus bacteria.  They reproduce by successive binary fission with a generation time of approximately 30 minutes with optimum growth occurring at 37 degrees centigrade.  E. coli are facilitative aerobic bacteria and are capable of ATP synthesis via both aerobic respiration and, if oxygen is not present, fermentation.  This particular strain can be identified and distinguished from other E. coli strains, by examining the genetic sequence of its 16s small ribosomal subunit, which has been fully sequenced (7).  
Ecology (including pathogenesis)
 
 
 
Ecology
 
This strain of E. coli was developed in the laboratory, for laboratory cloning procedures, and therefor has no 'natural' environment or habitat as it does not exist in nature. 
 


Describe its habitat, symbiosis, and contributions to environment. If it is a pathogen, how does this organism cause disease? Human, animal, plant hosts? Describe virulence factors and patient symptoms.
Interesting feature
Interesting feature


Describe in detail one particularly interesting aspect of your organism or it's affect on humans or the environment.
This specific strain of E. coli has been, in recent studies, used for computational data storage.  Researchers have been able to readily insert plasmids into the cell that contain computational data, encrypted using a quaternary coding language that makes use of the four nucleotide base pairs, into the cell and have it be self replicating (8).  This allows for excessive amounts of data storage, upwards of 900,000 gigabytes in a single gram of bacteria (9).  The research team, based in Hong Kong, have developed a computer program that corresponds with this research that allows users to 'simply' type, in regular text, what they would like to encode in the cell, and instantly have it translated into the biological code, which is then ready to be inserted into the cell (8).
 
References
References
(1) http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&id=668369&lvl=3&lin=f&keep=1&srchmode=1&unlock
(2) Taylor, R. G., Walker, D. C. and McInnes, R. R. ( 1993). E. coli host strains significantly affect the quality of small scale plasmid DNA preparations used for sequencing. Nucleic Acids Res. 21, 1677 -1678.
(3) http://bloodjournal.hematologylibrary.org/content/71/2/463.short
(4) http://www.ncbi.nlm.nih.gov/pubmed/18245285
(5) http://www.ncbi.nlm.nih.gov/pubmed/6363880
(6) http://www.ncbi.nlm.nih.gov/sites/entrez!db=genome&cmd=Retrieve&dopt=Overview&list_uids=22068
(7) Singh, Om V. ., and Steven P. . Harvey. "Bacterial Communities." Sustainable Biotechnology. Dordrecht [etc.: Springer, 2010.
169-71. Print.
(8) http://2010.igem.org/files/presentation/Hong_Kong-CUHK.pdf


[Sample reference] Takai, K., Sugai, A., Itoh, T., and Horikoshi, K. "Palaeococcus ferrophilus gen. nov., sp. nov., a barophilic, hyperthermophilic archaeon from a deep-sea hydrothermal vent chimney". International Journal of Systematic and Evolutionary Microbiology. 2000. Volume 50. p. 489-500.
(9) http://io9.com/5699767/bioencryption-can-store-almost-a-million-gigabytes-of-data-inside-bacteria

Latest revision as of 15:32, 17 December 2015

This student page has not been curated.

Classification

Domain: Bacteria Phylum: Protobacteria Class: Gammaproteobacteria Order: Enterobacteriales Family:Enterobacteriaceae Genus: Escherichia Species: Escherishia coli DH5-Alpha (1)


Description and significance

This strain of E. Coli is not a pathogen, and was developed for laboratory cloning use. This strain was developed by D. Hanahan as a cloning strain with multiple mutations that enable high-efficiency transformations (2). The mutations that the DH5-Alpha strain has are: dlacZ Delta M15 Delta(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 (2). These mutations correspond to the distinct characteristics that make the DH5-Alpha strain excel in laboratory cloning procedures (2).

lacZ Delta M15 mutation: Allows for blue-white screening for recombinant cells. (3) endA1 mutation: Allows for lower endonuclease degradation which ensures higher plasmid transfer rates. (4) recA1 mutation: reduces homologous recombination for a more stable insert. (5)


Genome structure

The genomic structure of this strain is a singular circular chromosome consisting of 4,686,137 nucleotides, 4359 genes, and 4128 protein encoding genes (6)

This strain also contains plasmids, and has the ability to accept plasmid insertion exceptionally well.


Cell structure, metabolism & life cycle

E. coli are gram-negative bacillus bacteria. They reproduce by successive binary fission with a generation time of approximately 30 minutes with optimum growth occurring at 37 degrees centigrade. E. coli are facilitative aerobic bacteria and are capable of ATP synthesis via both aerobic respiration and, if oxygen is not present, fermentation. This particular strain can be identified and distinguished from other E. coli strains, by examining the genetic sequence of its 16s small ribosomal subunit, which has been fully sequenced (7).


Ecology

This strain of E. coli was developed in the laboratory, for laboratory cloning procedures, and therefor has no 'natural' environment or habitat as it does not exist in nature.


Interesting feature

This specific strain of E. coli has been, in recent studies, used for computational data storage. Researchers have been able to readily insert plasmids into the cell that contain computational data, encrypted using a quaternary coding language that makes use of the four nucleotide base pairs, into the cell and have it be self replicating (8). This allows for excessive amounts of data storage, upwards of 900,000 gigabytes in a single gram of bacteria (9). The research team, based in Hong Kong, have developed a computer program that corresponds with this research that allows users to 'simply' type, in regular text, what they would like to encode in the cell, and instantly have it translated into the biological code, which is then ready to be inserted into the cell (8).

References (1) http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&id=668369&lvl=3&lin=f&keep=1&srchmode=1&unlock

(2) Taylor, R. G., Walker, D. C. and McInnes, R. R. ( 1993). E. coli host strains significantly affect the quality of small scale plasmid DNA preparations used for sequencing. Nucleic Acids Res. 21, 1677 -1678.

(3) http://bloodjournal.hematologylibrary.org/content/71/2/463.short

(4) http://www.ncbi.nlm.nih.gov/pubmed/18245285

(5) http://www.ncbi.nlm.nih.gov/pubmed/6363880

(6) http://www.ncbi.nlm.nih.gov/sites/entrez!db=genome&cmd=Retrieve&dopt=Overview&list_uids=22068

(7) Singh, Om V. ., and Steven P. . Harvey. "Bacterial Communities." Sustainable Biotechnology. Dordrecht [etc.: Springer, 2010. 169-71. Print.

(8) http://2010.igem.org/files/presentation/Hong_Kong-CUHK.pdf

(9) http://io9.com/5699767/bioencryption-can-store-almost-a-million-gigabytes-of-data-inside-bacteria