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Team:Stockholm/22 June 2010
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''Continued from 21/6''.<br /> | ''Continued from 21/6''.<br /> | ||
| - | Prepared competent Top10 cells according [https://2010.igem.org/wiki/index.php?title=Team:Stockholm/Protocols&action=submit#Competent_cells_.28Andreas.29 protocol]. | + | Prepared competent Top10 cells according to [https://2010.igem.org/wiki/index.php?title=Team:Stockholm/Protocols&action=submit#Competent_cells_.28Andreas.29 protocol]. |
=== Testing Top10 competent cells and antibiotic agar plates === | === Testing Top10 competent cells and antibiotic agar plates === | ||
Revision as of 10:05, 6 July 2010
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Contents |
Andreas
Preparation of competent Top10 Escherichia coli cells
Continued from 21/6.
Prepared competent Top10 cells according to protocol.
Testing Top10 competent cells and antibiotic agar plates
Tested competent Top10 by transforming with the BioBrick Freiburg expression vector pEX (BBa_K243033), carrying resistance to Amp, and plating onto previously prepared (21/6) 100 ug/ml Amp and 25 ug/ml Cm plates. Tested LB agar plates by plating with "mock-transformed" cells.
Hassan
- EuMelanin (EM)
- Melanin Gene in recombinant E. Coli.
- How IPTG effecting EM production.
- Study this article: http://www.ncbi.nlm.nih.gov/pubmed/17040223 Lagunas-Muñoz VH et. al. 2006 :
- Suggests that temperature and PH are the main factors effecting melanin production. they reached a 100% increase in melanin production by changing PH and Temp.
- The production of melanin is subjected to the presence of tyrosinase, which is an enzyme to catalyze the oxidation of phenols (for the melanin case this one is tyrosine) source: wikipedia.
- These enzymes have two activities: they use molecular oxygen to catalyse the hydroxylation of l-tyrosine to l-3,4- dihydroxyphenyl alanine (l-dopa) (cresolase activity); and then oxidizes this product to dopachrome (catecholase activity), which then non-enzymatically oxidizes and polymerizes to form melanin (Garcı´a-Borro´n and Solano 2002).
- EuMel production is what they were interested in, EuMel is directly related to induction of MutMelA gene, which could be controlled by concentration of IPTG. They suggest a 0.1 mmol/litr IPTG for mel production.
- Ask wetlab team for what their PH, Temp, and IPTG is for their bacteria.
- Possible pathway in EM production in E. Coli.
- What else is needed to represent the whole as system!?
- Overexpression / inclusion bodies
- TODO:
- Read about inclusion bodies, over expression.
- Read articles in production part of modeling and add more/delete some.
