Team:Hong Kong HKUST/Parts

• 
• Project
  Overview PE Degradation Alkane Metabolism Alkane Results MFC Design MFC Result Experimental protocol Notebook Parts
• Modelling
  Laccase Alkane Metabolism Alkane and MFC (FBA)
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  Education and Engagement Collaborations Integrated Human Practices
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Basic parts

BBa_K2764000

The gene alkL codes for the outer membrane alkane channel protein known as AlkL. Derived from Pseudomonas Oleovarans, the sequence of 693bp has already been codon optimized for an E.coli chassis. The AlkL is able to collect alkanes thanks to an extracellular domain that has a high binding affinity to hydrophobic molecules. Once collected, the alkane makes it way through the hydrophobic core of AlkL before exiting into the periplasm through a small lateral opening in the protein. Hence, the AlkL, which penetrates the lipopolysaccharide outer membrane, may allow alkanes of larger molecular size to enter the cell.
For more details, check out BBa_K2764000
From literature, we know that the AlkL protein is toxic. As such, we used Anderson promoters of different strength to determine which expression level would be optimal. The Anderson promoters selected were J23104, J23107 and J23110 were intended to form an expression level gradient for this purpose.

Composite Parts

BBa_K2764001

BBa_K2764001 utilizes J23104 as its promoter and has a relative promoter strength of 0.72 and stands at 891bp.
For more details, check out BBa_K2764001

BBa_K2764002

BBa_K2764002 utilizes J23108 as its promoter and has a relative promoter strength of 0.51 and stands at 891bp.
For more details, check out BBa_K2764002

BBa_K2764003

BBa_K2764002 utilizes J23110 as its promoter and has a relative promoter strength of 0.33 and stands at 891bp.
For more details, check out BBa_K2764003

REFERENCES:

1.Parts.igem.org. (2006). Promoters/Catalog/Anderson - parts.igem.org. [online] Available at: http://parts.igem.org/Promoters/Catalog/Anderson [Accessed 10 Sep. 2018].
2.Parts.igem.org. (2006). Part:BBa J23104 - parts.igem.org. [online] Available at: http://parts.igem.org/wiki/index.php/Part:BBa_J23104 [Accessed 10 Sep. 2018].
3.Parts.igem.org. (2006). Part:BBa J23108 - parts.igem.org. [online] Available at: http://parts.igem.org/wiki/index.php/Part:BBa_J23108 [Accessed 10 Sep. 2018].
4.Parts.igem.org. (2006). Part:BBa J23110 - parts.igem.org. [online] Available at: http://parts.igem.org/wiki/index.php/Part:BBa_J23110 [Accessed 10 Sep. 2018].
5.Grant, C., Deszcz, D., Wei, Y., Martínez-Torres, R., Morris, P., Folliard, T., Sreenivasan, R., Ward, J., Dalby, P., Woodley, J. and Baganz, F. (2014). Identification and use of an alkane transporter plug-in for applications in biocatalysis and whole-cell biosensing of alkanes.

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