Part:BBa_K1470004

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