Team:Cambridge/Project/ME01

From 2009.igem.org

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[https://2009.igem.org/Team:Cambridge/Project/ME01 Background]
[https://2009.igem.org/Team:Cambridge/Project/ME01 Background]
[https://2009.igem.org/Team:Cambridge/Project/ME02 Design]
[https://2009.igem.org/Team:Cambridge/Project/ME02 Design]
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[[Image:Tyrosinase action.jpg]]  
[[Image:Tyrosinase action.jpg]]  
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From H. Claus and H. Decker, Bacterial tyrosinases, Syst. Appl. Microbiol. 29 (2006), pp. 3–14. [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B7GVX-4H21K91-1&_user=1495569&_coverDate=01%2F24%2F2006&_fmt=full&_orig=search&_cdi=20442&view=c&_acct=C000053194&_version=1&_urlVersion=0&_userid=1495569&md5=de451c5e1d1d18ad12b7e39b10b80408&ref=full]
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From Claus and Decker, 2006
'''MelA'''
'''MelA'''
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Our MelA gene is from ''Rhizobium etli.'' Further, it is a mutant; it has a C to T substitution at the 1,000th nucleotide, which creates a Proline to Serine mutation that reduces the amount of time before melanin production is visible. (Santos, C. N., and G. Stephanopoulos. 2008. Melanin-based high-throughput screen for L-tyrosine production in Escherichia coli. Appl. Environ. Microbiol. 74:1190-1197 [http://aem.asm.org/cgi/reprint/74/4/1190]) The plasmid was provided by Christine Sanntos from the lab of G. Stephanopoulos to Duncan Rowe under the materials transfers agreement.
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Our MelA gene is from ''Rhizobium etli.'' Further, it is a mutant; it has a C to T substitution at the 1,000th nucleotide, which creates a Proline to Serine mutation that reduces the amount of time before melanin production is visible. (Santos et al. 2008) The plasmid pTRCmelA was provided by Christine Sanntos from the lab of G. Stephanopoulos to Duncan Rowe under the materials transfers agreement.
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=== Action plan of our team ===
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Using pTRCmelA as a template, we aim to show melanin expression in ''E. coli'' and isolate the MelA gene in biobrick form.
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Our action plan is as follows:
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:1. Test for melanin production
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:2. Isolate MelA gene in biobrick form
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:3. Integrate Mel biobrick into system (e.g amplification of logic gate system)
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Latest revision as of 01:27, 22 October 2009


Melanin Pigment

Background

Melanin Production

The MelA gene codes for a tyrosinase. Tyrosinases catalyze two reactions, as described in the figure below. Melanin is a macromolecular compound produced by the polymerization of the quinone product of the second reaction, and has a characteristic brown colour.

Tyrosinase action.jpg

From Claus and Decker, 2006

MelA

Our MelA gene is from Rhizobium etli. Further, it is a mutant; it has a C to T substitution at the 1,000th nucleotide, which creates a Proline to Serine mutation that reduces the amount of time before melanin production is visible. (Santos et al. 2008) The plasmid pTRCmelA was provided by Christine Sanntos from the lab of G. Stephanopoulos to Duncan Rowe under the materials transfers agreement.

Using pTRCmelA as a template, we aim to show melanin expression in E. coli and isolate the MelA gene in biobrick form.

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