Difference between revisions of "Part:BBa K1470004"

Revision as of 19:53, 16 October 2014

Secreted Alkaline Phosphatase (SEAP)

Template Secreted Alkaline Phosphatase

Usage and Biology

The Secreted Alkaline Phosphatase (SEAP) is an hydrolase enzyme which removes phosphate moieties from a wide range of molecules. It's build in the human placenta. A big advance is its auomatic excetrion into the extracellulare environment. This was achieved by a single amino acid mutation from L to R near the C-Terminus[Y]. SEAP is an ideal reporter protein for many applications. We used for an assay with para-Nitrophenylphosphate to measure the efficiency of our transfections and transductions by colour change and measurement of the absorption. Alternativly it's used to prevent DNA from self-ligating by removing the phosphate group or replace it with a radioactive version if radiolabeld DNA is need [X]. [x] Maxam AM, Gilbert W (1980). „Sequencing end-labeled DNA with base-specific chemical cleavages“. Meth. Enzymol. Methods in Enzymology 65 (1): 499–560. [Y] Lowe ME. „Site-specific mutations in the COOH-terminus of placental alkaline phosphatase: a single amino acid change converts a phosphatidylinositol-glycan-anchored protein to a secreted protein“. J Cell Biol. 1992 Feb;116(3):799-807.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 209
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 520
Illegal NgoMIV site found at 1489
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 850
Illegal BsaI.rc site found at 1339

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 __NOTOC__
 <partinfo>BBa_K1470004 short</partinfo>
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 Secreted Alkaline Phosphatase
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+ <p>The Secreted Alkaline Phosphatase (SEAP) is an hydrolase enzyme which removes phosphate moieties from a wide range of molecules. It's build in the human placenta. A big advance is its auomatic excetrion into the extracellulare environment. This was achieved by a single amino acid mutation from L to R near the C-Terminus[Y].
+SEAP is an ideal reporter protein for many applications. We used for an assay with para-Nitrophenylphosphate to measure the efficiency of our transfections and transductions by colour change and measurement of the absorption. Alternativly it's used to prevent DNA from self-ligating by removing the phosphate group or replace it with a radioactive version if radiolabeld DNA is need [X].
+[x] Maxam AM, Gilbert W (1980). „Sequencing end-labeled DNA with base-specific chemical cleavages“. Meth. Enzymol. Methods in Enzymology 65 (1): 499–560. [Y] Lowe ME. „Site-specific mutations in the COOH-terminus of placental alkaline phosphatase: a single amino acid change converts a phosphatidylinositol-glycan-anchored protein to a secreted protein“. J Cell Biol. 1992 Feb;116(3):799-807.
+</p>
 <!-- -->
 <span class='h3bb'>Sequence and Features</span>