Difference between revisions of "Part:BBa K2271060"

Revision as of 18:45, 29 October 2017

Snc1

This part is a truncated version of the v-SNARE (vesicle- synaptosome-associated-Soluble N-ethylmaleimide-sensitive-factor Attachment REceptorprotein) Snc1. Snc1 is in the wildtype form involved in the fusion of Golgi-derived secretory vesicles with the plasma membrane. Domains of the protein are a variable domain which is not important for the binding to the t-SNARE, H1 and H2 are the a-helical segments (forming the SNAREpin with the t-SNARE) and the transmembrane domain (Gerst Paper). For our approaches we used a truncated version without the transmembrane domain. This Snc1 truncation was fused to the N-Terminus of different peroxisomal membrane anchor (Link Pex15/Pex26) to secrete the compounds of this compartment (Bild?).

For testing this part we used a fusion with the N-Terminus of a Peroxisomal Membrane anchor. We co-expressed this construct with Gus-PTS to perform a GUS Assay.(oder: We performed a GUS-Assay by targetting GUS(beta-Glucuronidase) to the Peroxisome using a GUS-PTS1.) The secreted GUS in the supernatant was measured with the turnover of 4-methylumbelliferyl-beta-D-glucuronide to 4-methyl umbelliferone (4-MUG). The fluorescent 4-MUG was measured with a plate reader (excitation: 365 nm, emission: 465 nm).

Different peroxisomal membrane anchors were tested using the GUS-Assay. The highest activity of GUS could be measured in the supernatant of Pex15 (Link to part) as a Membrane Anchor.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2271060 short</partinfo>
-Vesicle membrane receptor protein (v-SNARE) that is involved in the fusion between Golgi-derived secretory vesicles with the plasma membrane.
+This part is a truncated version of the v-SNARE (vesicle- synaptosome-associated-Soluble N-ethylmaleimide-sensitive-factor Attachment REceptorprotein) Snc1. Snc1 is in the wildtype form involved in the fusion of Golgi-derived secretory vesicles with the plasma membrane. Domains of the protein are a variable domain which is not important for the binding to the t-SNARE, H1 and H2 are the a-helical segments (forming the SNAREpin with the t-SNARE) and the transmembrane domain (Gerst Paper). For our approaches we used a truncated version without the transmembrane domain. This Snc1 truncation was fused to the N-Terminus of different peroxisomal membrane anchor (Link Pex15/Pex26) to secrete the compounds of this compartment (Bild?).
-It can be used to be fused to a membrane anchor to secrete the target vesicle or compartment.
+For testing this part we used a fusion with the N-Terminus of a Peroxisomal Membrane anchor. We co-expressed this construct with Gus-PTS to perform a GUS Assay.(oder: We performed a GUS-Assay by targetting GUS(beta-Glucuronidase) to the Peroxisome using a GUS-PTS1.) The secreted GUS in the supernatant was measured with the turnover of 4-methylumbelliferyl-beta-D-glucuronide to 4-methyl umbelliferone (4-MUG). The fluorescent 4-MUG was measured with a plate reader (excitation: 365 nm, emission: 465 nm).
+Different peroxisomal membrane anchors were tested using the GUS-Assay. The highest activity of GUS could be measured in the supernatant of Pex15 (Link to part) as a Membrane Anchor.
 <!-- Add more about the biology of this part here