Purification and characterization of recombinant spider silk expressed in Escherichia coli

@article{Arcidiacono1998PurificationAC,
  title={Purification and characterization of recombinant spider silk expressed in Escherichia coli},
  author={Steven Arcidiacono and Charlene M. Mello and Drora Kaplan and Stephen Cheley and Hagan Bayley},
  journal={Applied Microbiology and Biotechnology},
  year={1998},
  volume={49},
  pages={31-38}
}
Abstract A partial cDNA clone, from the 3′ end of the dragline silk gene was isolated from Nephila clavipes major ampullate glands. This clone contains a 1.7-kb insert, consisting of a repetitive coding region of 1.4-kb and a 0.3-kb nonrepetitive coding region; 1.5-kb of the 1.7-kb fragment was cloned into Escherichia coli and a␣43-kDa recombinant silk protein was expressed. Characterization of the purified protein by Western blot, amino acid composition analysis, and matrix-assisted laser… 
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158 Citations
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158 Citations

Molecular cloning and expression of the C-terminus of spider flagelliform silk protein from Araneus ventricosus
TLDR
A cDNA coding for the C-terminus of spider flagelliform silk protein (AvFlag) was cloned from Araneus ventricosus and created a polyhedrin AvFlag fusion protein that is produced as recombinant polyhedra; this provides a basis for the source of spider silk proteins for various applications.
Cloning and Expression of Spider Dragline Silk Protein Gene in Escherichia coli and Eukaryotic Cells
TLDR
A beneficial attempt to develop the pathway for yielding spider silk protein by genetic engineering is provided.
Cloning and Expression of Spider Dragline Silk Protein Gene in Escherichia coli and Eukaryotic Cell
TLDR
A beneficial attempt to develop the pathway for yielding spider silk protein by genetic engineering is provided.
Spider silk-like proteins derived from transgenic Nicotiana tabacum
TLDR
The feasibility to produce recombinant proteins in transgenic tobacco Nicotiana tabacum with sequences representing spider silk protein building blocks with confirmed gene insertion and RNA transcription by PCR and reverse-transcriptase PCR is presented.
Construct Synthetic Gene Encoding Artificial Spider Dragline Silk Protein and its Expression in Milk of Transgenic Mice
TLDR
An artificial gene monomer, a 360 bp sequence, was designed and polymerized to encode an analog of dragline silk protein, which was successfully cloned into pBC1 vector and microinjected into the pronuclei of fertilized Kunming White eggs.
Molecular cloning and characterization of the partial major ampullate silk protein gene from the spider Araneus ventricosus
Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system
TLDR
The results showed it takes only 7 to 10 days to generate recombinant baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses.
Unique molecular architecture of egg case silk protein in a spider, Nephila clavata.
TLDR
A unique silk protein secreted from the cylindrical silk glands of the spider Nephila clavata, whose transcripts are homologous to one another in two termini and repetitive units, is described.
Stably Express Spider Flagelliform Silk Protein in Bombyx Mori Cell Line by PiggyBac Transposon-Derived Vector
TLDR
The present results suggested that transgenic silkworm directly secreting functional spider silk protein in cocoon by using piggyBac system is feasible.
Designing recombinant spider silk proteins to control assembly.
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References

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