Difference between revisions of "Part:BBa K1033914"

Revision as of 14:52, 13 October 2019

amajLime, yellow-green chromoprotein (incl RBS & J23110)

This chromoprotein from the coral Anemonia majano, amajLime (also known as amajCFP or amFP486), naturally exhibits strong color when expressed. The protein has an absorption maximum at 458 nm giving it a yellow-green color visible to the naked eye. Compared to many other chromoproteins, such as amilCP (BBa_K592009), amilGFP (BBa_K592010), spisPink (BBa_K1033932), asPink (BBa_K1033933) and aeBlue (BBa_K864401), the color development is slower. The color is readily observed in both LB or on agar plates after 24-48 hours of incubation. The protein amajLime has significant sequence homologies with proteins in the GFP family.

Characterisation

Team: Humboldt_Berlin 2019

ExPASy ProtParam Results:

Number of amino acids: 228

Molecular weight: 25185.49 Da

Theoretical pI: 7.27

Total number of negatively charged residues (Asp + Glu): 22

Total number of positively charged residues (Arg + Lys): 22

Extinction coefficients:

Ext. coefficient 26150 M-1 cm-1

Abs 0.1% (=1 g/l) 1.038, assuming all pairs of Cys residues form cystines

Ext. coefficient 25900 M-1 cm-1

Abs 0.1% (=1 g/l) 1.028, assuming all Cys residues are reduced

Instability index: The instability index (II) is computed to be 27.07. This classifies the protein as stable.

Aliphatic index: 51.71

Grand average of hydropathicity (GRAVY): -0.379

References

[http://www.nature.com/nbt/journal/v17/n10/full/nbt1099_969.html] Matz, Mikhail V., et al. "Fluorescent proteins from nonbioluminescent Anthozoa species." Nature biotechnology 17.10 (1999): 969-973.

[http://www.pnas.org/content/102/36/12712.short] Henderson, J. Nathan, and S. James Remington. "Crystal structures and mutational analysis of amFP486, a cyan fluorescent protein from Anemonia majano." Proceedings of the National Academy of Sciences of the United States of America 102.36 (2005): 12712-12717.

[http://www.ncbi.nlm.nih.gov/pubmed/18648549] Alieva, N. O., et al. 2008. Diversity and evolution of coral fluorescent proteins. PLoS One 3:e2680.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 5: / Line 5: @@
 ===Characterisation===
 Team: Humboldt_Berlin 2019
+----
+====ExPASy ProtParam Results:====
+'''Number of amino acids:''' 228
+'''Molecular weight:''' 25185.49 Da
+'''Theoretical pI:''' 7.27
+'''Total number of negatively charged residues (Asp + Glu):''' 22
+'''Total number of positively charged residues (Arg + Lys):''' 22
+'''Extinction coefficients:'''
+'''Ext. coefficient'''    26150 M-1 cm-1
+Abs 0.1% (=1 g/l)   1.038, assuming all pairs of Cys residues form cystines
+'''Ext. coefficient'''   25900 M-1 cm-1
+Abs 0.1% (=1 g/l)   1.028, assuming all Cys residues are reduced
+'''Instability index:''' The instability index (II) is computed to be 27.07. This classifies the protein as stable.
+'''Aliphatic index:''' 51.71
+'''Grand average of hydropathicity (GRAVY):''' -0.379
 ===References===
 [http://www.nature.com/nbt/journal/v17/n10/full/nbt1099_969.html] Matz, Mikhail V., et al. "Fluorescent proteins from nonbioluminescent Anthozoa species." Nature biotechnology 17.10 (1999): 969-973.