Abstract
A cDNA coding for the C-terminus of spider flagelliform silk protein (AvFlag) was cloned from Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvFlag consists of 167 amino acids of a repetitive region and 87 amino acids of a C-terminal non-repetitive region. The peptide motifs found in spider flagelliform silk proteins, GPGGX and GGX, were conserved in the repetitive region of AvFlag. Phylogenetic analysis further confirmed that AvFlag belongs to the spider flagelliform silk proteins. The AvFlag cDNA was expressed as a 28 kDa polypeptide in baculovirus-infected insect cells. As a new expression approach for spider silk protein, the combination of polyhedrin and AvFlag creates a polyhedrin AvFlag fusion protein (61 kDa) that is produced as recombinant polyhedra; this provides a basis for the source of spider silk proteins for various applications.
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Abbreviations
- BSA:
-
bovine serum albumin
- EST:
-
expressed sequence tag
- AvFlag:
-
flagelliform silk protein from A. ventricosus
- HRP:
-
horseradish peroxidase
- MOI:
-
multiplicity of infection
- PAUP:
-
Phylogenetic Analysis Using Parsimony
- PFU:
-
plaque-forming units
- PAGE:
-
polyacrylamide gel electrophoresis
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Lee, K.S., Kim, B.Y., Je, Y.H. et al. Molecular cloning and expression of the C-terminus of spider flagelliform silk protein from Araneus ventricosus . J Biosci 32, 705–712 (2007). https://doi.org/10.1007/s12038-007-0070-8
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DOI: https://doi.org/10.1007/s12038-007-0070-8