Difference between revisions of "Part:BBa K200013:Design"

Revision as of 15:33, 18 October 2009 (view source) JamesField (Talk | contribs) (→‎Design Notes) ← Older edit		Latest revision as of 15:37, 18 October 2009 (view source) JamesField (Talk | contribs) (→‎Cleavable Linker)
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	===Design Notes===		===Design Notes===
−	Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.
		+	==Cleavable Linker==
−	https://static.igem.org/mediawiki/2009/a/a2/Opiorphin.png	+	[[Image:Opiorphin.png|left|400px]]Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.
−		+
−	The methionine is shown in RED, the protease recognition site is shown in BLUE and opiorphin is shown in GREEN. The RED & BLUE sections make up the removable linker region cleaved by the serine protease enterokinase.	+
		+	The methionine is shown in RED, the protease recognition site is shown in BLUE and opiorphin is shown in GREEN. The RED & BLUE sections make up the removable linker region which is cleaved by the serine protease enterokinase.
		+	==Stop Codon==
	A double stop codon has been incorporated into the sequence to ensure efficient termination.		A double stop codon has been incorporated into the sequence to ensure efficient termination.
		+
		+
		+	==Codon Choice==
	The sequence has been optimised for expression in <i>E.coli</i>.		The sequence has been optimised for expression in <i>E.coli</i>.

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Latest revision as of 15:37, 18 October 2009

Opiorphin + EK cleavage site

Design Notes

Cleavable Linker

Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.

The methionine is shown in RED, the protease recognition site is shown in BLUE and opiorphin is shown in GREEN. The RED & BLUE sections make up the removable linker region which is cleaved by the serine protease enterokinase.

Stop Codon

A double stop codon has been incorporated into the sequence to ensure efficient termination.

Codon Choice

The sequence has been optimised for expression in E.coli.

Source

Synthetic part.

References