Latest revision as of 20:21, 27 October 2010

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Gram(-) Therapeutic
Gram(+) Therapeutic
Tools Created
- New Vectors
- New Software
Tools Used
- Software
- Next-Gen Cloning
Protocols
Safety
Meet the Team
Accomplishments
Parts Submitted
Notebook
References

Works Cited

Angelo Scorpio, Donald J. Chabot, William A. Day, David K. O'Brien, Nicholas J. Vietri, Yoshifumi Itoh, Mansour Mohamadzadeh, and Arthur M. Friedlander. Poly-gamma-Glutamate Capsule-Degrading Enzyme Treatment Enhances Phagocytosis and Killing of Encapsulated Bacillus anthracis Antimicrobial Agents and Chemotherapy, January 2007, p. 215-222,Vol. 51, No. 1. PMCID: PMC1797643

Angelo Scorpio, Donald J Chabot, William A Day, Timothy A Hoover, and Arthur M Friedlander Capsule depolymerase overexpression reduces Bacillus anthracis virulence Microbiology 2010 : mic.0.035857-0v1-mic.0.035857-0.

Candela T, Fouet A. Bacillus anthracis CapD, belonging to the gamma-glutamyltranspeptidase family, is required for the covalent anchoring of capsule to peptidoglycan. Mol Microbiol. 2005 Aug;57(3):717-26. PMID: 16045616

Chandran D. [http://www.jbioleng.org/content/3/1/19 TinkerCell: modular CAD tool for synthetic biology]. Journal of Biological Engineering 2009, 3:19.

Filloux, Alain, Hachani, Abderrahman, Bleves, Sophie [http://mic.sgmjournals.org/cgi/content/full/154/6/1570#SEC7 The bacterial type VI secretion machine: yet another player for protein transport across membranes] Microbiology 2008 154: 1570-1583

Hood RD, Singh P, Hsu F, Güvener T, Carl MA, Trinidad RR, Silverman JM, Ohlson BB, Hicks KG, Plemel RL, Li M, Schwarz S, Wang WY, Merz AJ, Goodlett DR, Mougous JD. [http://www.cell.com/cell-host-microbe/abstract/S1931-3128%2809%2900417-X#Summary A type VI secretion system of Pseudomonas aeruginosa targets a toxin to bacteria]. Cell Host Microbe. 2010 Jan 21;7(1):25-37. PMID: 20114026

T.A. KUNKEL, P NATL ACAD SCI USA 82, 488 (JANUARY 1985, 1985)

Lucey DR, Anthrax. In: Goldman L, Ausiello D, eds. Cecil Medicine. 23rd ed. Philadelphia, Pa: Saunders Elsevier; 2007:chap 317.

Wu R, Richter S, Zhang RG, Anderson VJ, Missiakas D, Joachimiak A. J Biol Chem. 2009 Sep 4;284(36):24406-14. Epub 2009 Jun 16. PMID: 19535342

← Lab Notebook

@@ Line 25: / Line 25: @@
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+=Works Cited=
-'''Include references'''
+Angelo Scorpio, Donald J. Chabot, William A. Day, David K. O'Brien, Nicholas J. Vietri, Yoshifumi Itoh, Mansour Mohamadzadeh, and Arthur M. Friedlander.  Poly-gamma-Glutamate Capsule-Degrading Enzyme Treatment Enhances Phagocytosis and Killing of Encapsulated Bacillus anthracis Antimicrobial Agents and Chemotherapy, January 2007, p. 215-222,Vol. 51, No. 1.  PMCID: PMC1797643
-[[Image:CapDCP_MassSpecNoMethionine.png|thumb|780px|right| Expected weight of CapD_CP without Methionine=55285Da, with Methionine=55417Da. Our mass spec detected a peak at 55274.8Da (no Methionine) well within the 0.02% error limit for our mass spec]]
+Angelo Scorpio, Donald J Chabot, William A Day, Timothy A Hoover, and Arthur M Friedlander
+Capsule depolymerase overexpression reduces Bacillus anthracis virulence
+Microbiology 2010 : mic.0.035857-0v1-mic.0.035857-0.
-=Using Recombineering to create a constitutive T6SS=
+Candela T, Fouet A.  Bacillus anthracis CapD, belonging to the gamma-glutamyltranspeptidase family, is required for the covalent anchoring of capsule to peptidoglycan.
-[[Image:Washington Recombineering.jpg|400px|thumb|right|Insertion of T7 promoters]]
+Mol Microbiol. 2005 Aug;57(3):717-26.  PMID: 16045616
-The process for using homologous recombination to replace the T6SS native promoters is simple.  We begin with a previously characterised fosmid (a large, single-copy plasmid based on the F' plasmid) which contains all the genes in our T6SS.  Of particular interest is the ''fha1-tssA1'' intergenic region containing the promoters.  We transform this fosmid into recombineering strain ''E. coli'' SW102, which contains heat-inducible lambda phage recombinases and lacks the galactose metabolism gene ''galK''.
-Using PCR, we create a cassette containing the ''galK'' gene flanked by 50 bp of homology to the intergenic region.  The lambda recombinases integrate the cassette into the fosmid.  The transformants are plated on minimal galactose media to select for the recombinants.
+Chandran D. [http://www.jbioleng.org/content/3/1/19 TinkerCell: modular CAD tool for synthetic biology].
+Journal of Biological Engineering 2009, 3:19.
-A second cassette is created via PCR, this time containing our new promoters.  In this case, we chose to use T7 phage promoters due to their constitutive expression and small size, making them easier to synthesize.  The promoters are again flanked with homologous regions, and integrated into the fosmid.  This cassette displaces the ''galK'' cassette.  The transformants are this time plated on 2-deoxy-galactose (DOG), which is a toxic homolog to galactose.  Cells which can metabolize galactose (that is, those with a functioning ''galK'' gene) are killed, while those without the gene are unharmed.
+Filloux, Alain, Hachani, Abderrahman, Bleves, Sophie
+[http://mic.sgmjournals.org/cgi/content/full/154/6/1570#SEC7 The bacterial type VI secretion machine: yet another player for protein transport across membranes]
+Microbiology 2008 154: 1570-1583
-The final step is to transform the recombinant plasmid into a T7 polymerase expression strain, to allow T6SS constitutive expression.
+Hood RD, Singh P, Hsu F, Güvener T, Carl MA, Trinidad RR, Silverman JM, Ohlson BB, Hicks KG, Plemel RL, Li M, Schwarz S, Wang WY, Merz AJ, Goodlett DR, Mougous JD. [http://www.cell.com/cell-host-microbe/abstract/S1931-3128%2809%2900417-X#Summary A type VI secretion system of Pseudomonas aeruginosa targets a toxin to bacteria].
+Cell Host Microbe. 2010 Jan 21;7(1):25-37. PMID: 20114026
+T.A. KUNKEL, P NATL ACAD SCI USA 82, 488 (JANUARY 1985, 1985)
+Lucey DR, Anthrax. In: Goldman L, Ausiello D, eds. Cecil Medicine. 23rd ed. Philadelphia, Pa: Saunders Elsevier; 2007:chap 317.
+Wu R, Richter S, Zhang RG, Anderson VJ, Missiakas D, Joachimiak A. J Biol Chem. 2009 Sep 4;284(36):24406-14. Epub 2009 Jun 16. PMID: 19535342
+[[Image: Washington Get away.jpg]]
 <!---------------------------------------PAGE CONTENT GOES ABOVE THIS---------------------------------------->
 <div style="text-align:center">
-'''&larr; [[Team:Washington/Accomplishments|Accomplishments and Submitted Parts]] &rarr;'''
+'''&larr; [[Team:Washington/Notebook|Lab Notebook]]'''
 </div>
 {{Template:Team:Washington/Templates/Footer}}

Team:Washington/Bibliography

From 2010.igem.org

Latest revision as of 20:21, 27 October 2010

Works Cited