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

Revision as of 14:43, 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)
(6 intermediate revisions by the same user not shown)
Line 6:		Line 6:
	===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==
		+
		+	[[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 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>.

---

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