Difference between revisions of "Part:BBa K2235007"

Revision as of 13:13, 18 October 2017

OmpR promoter regulating the expression of sialidase enzyme

OmpR is an osmotic pressure promoter. It can sense changes in for example sucrose concentration. A higher concentration of sucrose will lead to a higher expression of the gene after the OmpR promoter. In this biobrick the gene after OmpR is an enzyme called sialidase. This enzyme cuts sialic acid (that's for example found in mucus). The sialidase produced by this biobrick has a HIS-tag on it to make it easy to purify.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 189
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 636
Illegal NgoMIV site found at 711
Illegal NgoMIV site found at 801
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1181

Usage and biology

OmpR is an already well characterized biobrick (BBa_R0082). It is a promoter that takes advantage of the OmpR/EnvZ two-component system, which is utilized by many bacteria, to sense the change in osmotic pressure. We connected this promoter to the coding sequence for the enzyme sialidase. This enzyme hydrolyzes glycosidic linkages of terminal sialic acid residues in glycoproteins. This composite biobrick can be used to produce sialidase when the osmotic pressure in the environment around the bacteria increases.

Important parameters

Experiment	Characteristic	Result
Expression
	Compatibility	E. coli TOP10 E. coli ΔEnvZ
	Promoter	OmpR
	Optimal temperature	37 °C
	Number of amino acids	XXX
	Molecular weight	XXXnbsp;kDa
Purification
Purification	Tag	C-terminal 6xHis
Sialidase expression
Sialidase expression	SDS-PAGE	Sialidase expression seen in some tests
Osmotic pressure detection
	Sucrose	Works
	NaCl	Non-conclusive data

@@ Line 60: / Line 60: @@
 |Sucrose
 |Works
+|-
 |NaCl
 |Non-conclusive data
 |}
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