Difference between revisions of "Part:BBa K1499400"

Revision as of 00:33, 25 October 2014

Chitin-binding wasp protein 1 (PdomMRNAr1.2-03231.1)

For more information on this project, visit our wiki: http://2014.igem.org/Team:StanfordBrownSpelman/Material_Waterproofing

Usage and Biology

Paper wasps of the genus Polistes are well known for their ability to construct nests composed of a tough, fibrous paper-like material. Studies have shown that the wasps build these nests by collecting cellulose from plants in the environment, chewing it up, and mixing with a protein-rich oral secretion that helps strengthen and cement the cellulose into nest paper[1]. Interestingly, these nests are also hydrophobic, which leads us to believe there may be a single protein in wasp saliva responsible for coating and waterproofing the nest paper.

In an attempt to identify and characterize this protein, we collected samples of nests constructed by Polistes dominula and extracted total protein. After resolving the protein extract on polyacrylamide gels, we excised dominant bands and sent them to the Wessel lab at Brown University for peptide mass fingerprinting.

Figure 1. Wasp nest protein extracts run on a polyacrylamide gel after a ten minute 70ºC denaturation step.

Figure 2. Wasp nest protein extracts run on a polyacrylamide gel without any denaturation step.

After analyzing the peptide mass fingerprinting results, we selected this protein as a candidate for the hypothesized waterproofing protein, due to PSI-BLAST results indicating that it may have a chitin-binding domain (chitin is a close chemical relative of cellulose).

The full suite of proteins that we investigated, along with additional information about this protein is viewable in the following table (Figure 3).

Figure 3. Wasp candidate genes.

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
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 143
1000
COMPATIBLE WITH RFC[1000]

References

1. Kudo, K et al. (2000) Amino acid composition of the protein in pre-emergence nests of a paper wasp, Polistes chinensis (Hymenoptera, Vespidae). Insectes soc. 47(4):371-375.

@@ Line 8: / Line 8: @@
 Paper wasps of the genus <i>Polistes</i> are well known for their ability to construct nests composed of a tough, fibrous paper-like material. Studies have shown that the wasps build these nests by collecting cellulose from plants in the environment, chewing it up, and mixing with a protein-rich oral secretion that helps strengthen and cement the cellulose into nest paper[1]. Interestingly, these nests are also hydrophobic, which leads us to believe there may be a single protein in wasp saliva responsible for coating and waterproofing the nest paper.
-In an attempt to identify and characterize this protein, we collected samples of nests constructed by <i>Polistes dominula</i> and extracted total protein. After resolving the protein extract on a polyacrylamide gel, we excised dominant bands and sent them to the Wessel lab at Brown University for peptide mass fingerprinting.
+In an attempt to identify and characterize this protein, we collected samples of nests constructed by <i>Polistes dominula</i> and extracted total protein. After resolving the protein extract on polyacrylamide gels, we excised dominant bands and sent them to the Wessel lab at Brown University for peptide mass fingerprinting.
 [[Image:SBS_iGEM_Ian_Hull_Wasp_Nest_Gel_Final.png|750px|thumb|center|<b>Figure 1.</b> Wasp nest protein extracts run on a polyacrylamide gel after a ten minute 70ºC denaturation step.]]