IGEM:Harvard/2010/Brainstorming/food: Difference between revisions
No edit summary |
|||
Line 14: | Line 14: | ||
===protocols=== | ===protocols=== | ||
====Growing a Bacterial E. coli Culture==== | |||
====Plasmid Minipreps: Qiagen Miniprep Kit==== | |||
====DNA Digests==== | |||
====DNA Gel Electrophoresis and Fragment Purification==== | |||
====Measuring DNA Concentration using a Nanodrop Spectrophotometer==== | |||
====Ligation Reaction==== | |||
====Transformation of E. coli==== | |||
====Confirming the Assembly==== | |||
*Growing Arabidopsis | *Growing Arabidopsis |
Revision as of 10:20, 17 May 2010
Strains, Plasmids, and Trasformation
references
1. Broothearts et. al. Gene transfer to plants by diverse species of bacteria. Nature, 2004.
2. Galvin [1] Agrobacterium-Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool
Agrobacterium infection diagram
3. Antunes, et. al. [2] A synthetic de-greening gene circuit provides a reporting system that is remotely detectable and has a re-set capacity
4. Everything you want to know about Arabidopsis [3]
protocols
Growing a Bacterial E. coli Culture
Plasmid Minipreps: Qiagen Miniprep Kit
DNA Digests
DNA Gel Electrophoresis and Fragment Purification
Measuring DNA Concentration using a Nanodrop Spectrophotometer
Ligation Reaction
Transformation of E. coli
Confirming the Assembly
- Growing Arabidopsis
- Ti Plasmids
- Growing an Agrobacterium Culture
- Arabidopsis transformation via Flower Dipping
- Book: Agrobacterium Protocols
RNAi
- Chuang CF and Meyerowitz EM. Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4985-90. DOI:10.1073/pnas.060034297 |
Markers and Genetic Fences
Dual selection system ideas
- use selectable markers such as URA3 to control whether an integration plasmid is replicated or not
- cells with the URA3 gene can grow on media without uracil, but get killed by 5-Fluoroorotic acid (5-FOA)
- cells without the URA3 gene die on media without uracil, but can survive exposure to 5-FOA
De-Greening
- this paper has some parts that could be useful as kill-switches
- A synthetic de-greening gene circuit provides a reporting system that is remotely detectable and has a re-set capacity.
- steroid-inducible chlorophyll knockdown
Downstream effectors
allergens
Peanut and Tomatoes review
- Singh MB and Bhalla PL. Genetic engineering for removing food allergens from plants. Trends Plant Sci. 2008 Jun;13(6):257-60. DOI:10.1016/j.tplants.2008.04.004 |
Peanut profilins: Ara h knock-down
- Dodo H, Konan K, and Viquez O. A genetic engineering strategy to eliminate peanut allergy. Curr Allergy Asthma Rep. 2005 Jan;5(1):67-73. DOI:10.1007/s11882-005-0058-0 |
Tomato profilins: Lyc e knock-down
- Design of tomato fruits with reduced allergenicity by dsRNAi-mediated inhibition of ns-LTP (Lyc e 3) expression. Le LQ, Lorenz Y, Scheurer S, Fötisch K, Enrique E, Bartra J, Biemelt S, Vieths S, Sonnewald U. Plant Biotechnol J. 2006 Mar;4(2):231-42. pmid:17177799
- Reduced allergenicity of tomato fruits harvested from Lyc e 1-silenced transgenic tomato plants. Le LQ, Mahler V, Lorenz Y, Scheurer S, Biemelt S, Vieths S, Sonnewald U. J Allergy Clin Immunol. 2006 Nov;118(5):1176-83. Epub 2006 Sep 8. pmid:17088146
Orange profilin: Cit s 2 (characterized, but not knocked down yet)
- Isolation, cloning and allergenic reactivity of natural profilin Cit s 2, a major orange allergen. López-Torrejón G, Ibáñez MD, Ahrazem O, Sánchez-Monge R, Sastre J, Lombardero M, Barber D, Salcedo G. Allergy. 2005 Nov;60(11):1424-9. pmid:16197476
flavors
- [4] Tieman et al. Tomato aromatic amino acid decarboxylases participate in synthesis of the flavor volatiles 2-phenylethanol and 2-phenylacetaldehyde
VANILLA: http://www.3dchem.com/moremolecules.asp?ID=307&othername=Vanilla#
- Sinha AK, Sharma UK, and Sharma N. A comprehensive review on vanilla flavor: extraction, isolation and quantification of vanillin and others constituents. Int J Food Sci Nutr. 2008 Jun;59(4):299-326. DOI:10.1080/09687630701539350 |
De novo biosynthesis of vanillin in fission yeast (Schizosaccharomyces pombe) and baker's yeast (Saccharomyces cerevisiae). http://www.ncbi.nlm.nih.gov.ezp-prod1.hul.harvard.edu/pubmed/19286778
miraculin
spices
Capsaicin: http://en.wikipedia.org/wiki/Capsaicin
- Stewart C Jr, Kang BC, Liu K, Mazourek M, Moore SL, Yoo EY, Kim BD, Paran I, and Jahn MM. The Pun1 gene for pungency in pepper encodes a putative acyltransferase. Plant J. 2005 Jun;42(5):675-88. DOI:10.1111/j.1365-313X.2005.02410.x |
- Prasad BC, Kumar V, Gururaj HB, Parimalan R, Giridhar P, and Ravishankar GA. Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicum sp.). Proc Natl Acad Sci U S A. 2006 Sep 5;103(36):13315-20. DOI:10.1073/pnas.0605805103 |
From wikipedia:
- TRPV1, the only known receptor (a transient receptor potential channel) for capsaicin.
- Piperine, the active piquant chemical in black pepper
- Allyl isothiocyanate, the active piquant chemical in mustard, radishes, horseradish, and wasabi
- Allicin, the active piquant flavor chemical in uncooked garlic and onions (see those articles for discussion of other chemicals in them relating to pungency, and eye irritation)
- Naga Jolokia pepper, the world's most capsaicin-rich fruit
- Discovery and development of TRPV1 antagonists
- Capsinoids are similar in structure to capsaicin, but lack the extreme pungency.
others
- Raksakulthai R and Haard NF. Exopeptidases and their application to reduce bitterness in food: a review. Crit Rev Food Sci Nutr. 2003;43(4):401-45. DOI:10.1080/10408690390826572 |
http://en.wikipedia.org/wiki/Limonene
caffeine etc.
- Ashihara H, Sano H, and Crozier A. Caffeine and related purine alkaloids: biosynthesis, catabolism, function and genetic engineering. Phytochemistry. 2008 Feb;69(4):841-56. DOI:10.1016/j.phytochem.2007.10.029 |
Questions
can we eat it for the iGEM project (human testing)? Can we hand it out to be eaten (FDA)? Patent/copyright issues? Can we have actual tranformed plants ready by the jamboree