Team:Sharon MA Aquila/Project

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{{Aquila}}
{{Aquila}}
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The purpose of our project is to insert antifreeze protein (AFP) into yogurt cultures.  Milk is converted to yogurt using  ''Lactobacillus delbrueckii'' subsp. ''bulgaricus'' and ''Streptococcus salivarius'' subsp. ''thermophilus'' bacteria.  These cultures ferment the lactose sugar present in the milk and convert it to lactic acid, which gives yogurt its distinctive taste and texture.
 
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AFP is a protein produced by plants, fungi, bacteria, and some vertebrates to prevent the formation of dangerous ice crystals at subzero temperatures. AFP is about 300 times more effective than industrially produced antifreezes at the same concentration, so it hasmany possible commercial applications, including cryosurgery, hypothermia treatment, and farm fish production.  Some companies have also started isolating AFPs from fish and introducing them into milk and yogurt products.  The protein could help prevent freezer burn and improve the texture of the yogurt (frozen or otherwise).
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===Parts===
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[http://http://partsregistry.org/Part:BBa_K116001 Our pH sensor]
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Our project will instead introduce the AFP-producing gene directly into the ''Lactobacillus bulgaricus'' bacteria, so that it is constantly produced as the bacteria metabolizes lactose. Hooking up production of the protein to the lactose sensor already present in the bacteria would allow continuous production of the protein. Since transplantation of a small quantity of yogurt cultures can be used to produce another batch of yogurt, our altered bacteria would easily be able to replicate and produce AFP in new batches.
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===Materials===
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<br>
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*'''Culturing:'''
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**[http://www.bd.com/ds/productCenter/288110.asp MRS broth]
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**[http://www.bd.com/ds/productCenter/288210.asp MRS agar]
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*'''Bacterial Transformation:'''
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**Electroporation Buffer:(0.4 M sucrose, 1 mM MgCl2, 5 mM Kh2PO4; PH 6)
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**Erythromycin (for selection)
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**Gene Pulser and Pulse Controller Electrophoration apparatus
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**Gaspak or method to generate anaerobic conditions (airtight box with lit candles)
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*'''Miniprep'''
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**TES(50mM Tris[pH 7], 20mM EDTA, 0.2 mM sucrose)
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**10mg/ml lysozyme
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**70U/ml mutanolysin
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'''Parts:'''
 
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*[http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=11842&Template=bacteria ''Lactobacillus delbrueckii'' subsp. ''bulgaricus'']
 
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*[http://partsregistry.org/Part:BBa_K652001 RiAFP part] (from the Parts Registry)
 
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*MRS broth (for culturing)
 
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*MRS agar (for culturing)
 
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Latest revision as of 19:37, 21 May 2013


Parts

[http://http://partsregistry.org/Part:BBa_K116001 Our pH sensor]

Materials

  • Culturing:
    • [http://www.bd.com/ds/productCenter/288110.asp MRS broth]
    • [http://www.bd.com/ds/productCenter/288210.asp MRS agar]
  • Bacterial Transformation:
    • Electroporation Buffer:(0.4 M sucrose, 1 mM MgCl2, 5 mM Kh2PO4; PH 6)
    • Erythromycin (for selection)
    • Gene Pulser and Pulse Controller Electrophoration apparatus
    • Gaspak or method to generate anaerobic conditions (airtight box with lit candles)
  • Miniprep
    • TES(50mM Tris[pH 7], 20mM EDTA, 0.2 mM sucrose)
    • 10mg/ml lysozyme
    • 70U/ml mutanolysin


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