Difference between revisions of "Part:BBa K1033932"

Latest revision as of 13:35, 12 October 2019

spisPink, pink chromoprotein

This chromoprotein from the coral Stylophora pistillata, spisPink (also known as spisCP), naturally exhibits strong color when expressed. The protein has an absorption maximum at 560 nm giving it a pink color visible to the naked eye. The strong color is readily observed in both LB or on agar plates after less than 24 hours of incubation. The protein spisPink has significant sequence homologies with proteins in the GFP family.

Important: This part is not available in the registry yet, however, the same part is available from the registry with the standard RBS B0034: BBa_K1033925.

Usage and Biology

This part is useful as a reporter.

spisPink does not exist as CDS only but only as RBS-spisPink (BBa_K1033925), J23110-RBS-spisPink (BBa_K1033923) and J23106-RBS-spisPink (BBa_K1033924)

iGEM2013 Uppsala: The images above show E coli constitutively expressing spisPink BBa_K1033925 from the high copy plasmid pSB1C3 from the promoters J23110 (left) and J23106 (right).

Source

Stylophora pistillata. The protein was first extracted and characterized by Alieva et. al. under the name spisCP (GenBank: ABB17971.1). This version is codon optimized for E coli by Genscript.

References

[http://www.ncbi.nlm.nih.gov/pubmed/18648549] Alieva, Naila O., et al. "Diversity and evolution of coral fluorescent proteins." PLoS One 3.7 (2008): e2680.

Contribution

Group: Team Tsinghua 2017

Author: Boyang Gao

Summary: We have improved the characterization of this part by constructing it into a yeast expression vector and expressing it in a new chassis organism, S. cerevisiae.

The expression of spisPink protein in yeast cells

Yeast cells were transformed with plasmids (backbone: pRS424; Promoter: TEF2; Terminator: CYC1) constitutively expressing spisPink protein. auxotrophic medium was used to ensure the successful transformation of plasmids, and this picture was taken 72 h after transformation. As shown in the picture, the yeast colonies did not turn pink apparently. Thus spisPink might not work as well in yeast cells as in bacteria.

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]

Characterization: NWU-China 2019

The NWU-China team overexpressed spisPink protein in E. coli and extracted and purified. The maximum absorbance was measured in a 96-well plate using a microplate reader (Synergy 2).

Figure 1. spisPink absorbance spectrum

@@ Line 3: / Line 3: @@
 This chromoprotein from the coral ''Stylophora pistillata'', spisPink (also known as spisCP), naturally exhibits strong color when expressed. The protein has an absorption maximum at 560 nm giving it a pink color visible to the naked eye. The strong color is readily observed in both LB or on agar plates after less than 24 hours of incubation. The protein spisPink has significant sequence homologies with proteins in the GFP family.
+'''Important: This part is not available in the registry yet, however, the same part is available from the registry with the standard RBS B0034: <partinfo>BBa_K1033925</partinfo>.'''
 ===Usage and Biology===
 This part is useful as a reporter.
+spisPink does not exist as CDS only but only as RBS-spisPink (<partinfo>BBa_K1033925</partinfo>), J23110-RBS-spisPink (<partinfo>BBa_K1033923</partinfo>) and J23106-RBS-spisPink (<partinfo>BBa_K1033924</partinfo>)
 [[Image:SpisPink1.jpg|500px]]
-'''iGEM2013 Uppsala:''' The images above show ''E coli'' constitutively expressing spisPink <partinfo>BBa_K1033932</partinfo> from the high copy plasmid pSB1C3 from the promoters J23110 (left) and J23106 (right).
+'''iGEM2013 Uppsala:''' The images above show ''E coli'' constitutively expressing spisPink <partinfo>BBa_K1033925</partinfo> from the high copy plasmid pSB1C3 from the promoters J23110 (left) and J23106 (right).
 ===Source===
@@ Line 16: / Line 22: @@
 ===References===
 [http://www.ncbi.nlm.nih.gov/pubmed/18648549] Alieva, Naila O., et al. "Diversity and evolution of coral fluorescent proteins." PLoS One 3.7 (2008): e2680.
+=====Contribution=====
+Group: Team Tsinghua 2017
+Author: Boyang Gao
+Summary: We have improved the characterization of this part by constructing it into a yeast expression vector and expressing it in a new chassis organism, S. cerevisiae.
+[[File:SpisPink expression in yeast cells.jpeg|500px]]
+=====The expression of spisPink protein in yeast cells=====
+Yeast cells were transformed with plasmids (backbone: pRS424; Promoter: TEF2; Terminator: CYC1) constitutively expressing spisPink protein. auxotrophic medium was used to ensure the successful transformation of plasmids, and this picture was taken 72 h after transformation. As shown in the picture, the yeast colonies did not turn pink apparently. Thus spisPink might not work as well in yeast cells as in bacteria.
@@ Line 27: / Line 50: @@
 <partinfo>BBa_K1033932 parameters</partinfo>
 <!-- -->
+===Characterization: NWU-China 2019===
+The NWU-China team overexpressed spisPink protein in E. coli and extracted and purified. The maximum absorbance was measured in a 96-well plate using a microplate reader (Synergy 2).
+<br>
+[[File:T--NWU-China--characterization-BBa_K1033932%28spisPink%29.png|900px|thumb|none|alt=fwYellow spectum.|Figure 1. spisPink absorbance spectrum]]
+;