Production and purification of recombinant DP1B silk-like protein in plants

Barr, Leslie A; Fahnestock, Stephen R.; Yang, Jianjun

doi:10.1023/b:molb.0000034089.92263.ff

Cited by 41 publications

(42 citation statements)

References 18 publications

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“…The recombinant silk protein could easily be purified by a combination of heat treatment, acidification and ammonium sulfate precipitation [35,41]. In addition to tobacco and potatoes another engineered spidroin based on MaSp1 from Nephila clavipes was produced in the seeds of Arabidopsis, yielding 18% of the desired engineered spidroin within the total soluble protein, which makes this a viable means of protein production [42,43].…”

Section: Recombinant Production Of Engineered Spidroins With Adapted mentioning

confidence: 99%

Recombinant Spider Silks—Biopolymers with Potential for Future Applications

2011

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Nature has evolved a range of materials that compete with man-made materials in physical properties; one of these is spider silk. Silk is a fibrous material that exhibits extremely high strength and toughness with regard to its low density. In this review we discuss the molecular structure of spider silk and how this understanding has allowed the development of recombinant silk proteins that mimic the properties of natural spider silks. Additionally, we will explore the material morphologies and the applications of these proteins. Finally, we will look at attempts to combine the silk structure with chemical polymers and how the structure of silk has inspired the engineering of novel polymers.

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Section: Recombinant Production Of Engineered Spidroins With Adapted mentioning

confidence: 99%

Recombinant Spider Silks—Biopolymers with Potential for Future Applications

2011

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“…The oligonucleotides can be ligated to constitute silk genes expressible in bacteria ( Figure 2 and see below). [18][19][20][21][22][23][24][25][26][27][28][29][30][31][32] Another approach is based on expressing authentic spider cDNA in optimized hosts other than bacteria that can be produced at large scale. Furthermore, several eukaryotic hosts are able to ''decode'' the codon usage of spider genes, are not size-limited in gene expression, and have a low frequency of gene recombination.…”

Section: Why Is Spider Silk a Prime Target For Biomaterials Researchers?mentioning

confidence: 99%

“…Plants like tobacco, potato and Arabidopsis thaliana have further been employed as transgenic expression hosts for synthetic silk genes, grown in both greenhouse and field trials ( Table 3). [30][31][32][47][48][49] Therein, the silk proteins have been targeted to different compartments in order to gain high yields. In tobacco leaves a yield of 2% (w/w) of total protein was reached when targeted to the endoplasmic reticulum.…”

Section: Production Of Engineered Silk Proteinsmentioning

confidence: 99%

Biotechnological Production of Spider‐Silk Proteins Enables New Applications

Vendrely

Scheibel

2007

Macromolecular Bioscience

220

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The outstanding mechanical properties of spider silks have motivated many researchers to establish biotechnological production techniques which are necessary to provide sufficient amounts of silk proteins for industrial applications. Based on recent developments in genetic engineering, two strategies for the recombinant production of spider-silk proteins have been established which are discussed in detail. Further, protein-design strategies are described, enabling the combination of silk properties with additional biological, chemical, or technical features. We highlight the potential of engineered and recombinantly-produced spider-silk proteins to provide the basis for a new generation of biomaterials.

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“…Des Weiteren werden wäh-rend der Induktion die tRNA-und Aminosäurevorräte der Zelle wegen des überproportional hohen Anteils von Glycin, Glutamin, Alanin, Prolin und Serin schnell aufgebraucht (Abbildung 2 a). Um die genannten Schwierigkeiten zu umgehen, wurden Spinnenseidenproteine in genetisch verän-derten Bakterien, [38][39][40][41][42] Hefen, [40,43] Pflanzen, [44][45][46] Insekten- [47] und Säugerzellen [48] sowie in transgenen Tieren [49] hergestellt. Jeder dieser Wirtsorganismen bietet gewisse Vor-, aber auch Nachteile (Tabelle 3).…”

Section: Die Biotechnologische Herstellung Von Spinnenseideunclassified

“…b) Vergleich dieser fünf Proteine hinsichtlich der gesamten Aminosäurezusammensetzung, gegliedert nach ihren chemischen Charakteristika, basierend auf den publizierten Sequenzen: U47855 (ADF-3), NP 001092 (b-Aktin), NP 851335 (BSA), P69905 (Hämoglobin-a-Untereinheit) und NM 000088 (Kollagen Typ I, a1). Pflanzen: Arabidopsis thaliana [44] Solanum tuberosum (Kartoffel) [45,46] Nicotiana (Tabak) [45,46] MAS und abgeleitete Proteine …”

Section: Die Biotechnologische Herstellung Von Spinnenseideunclassified

Spinnenseide: vom löslichen Protein zur außergewöhnlichen Faser

2009

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Am seidenen Faden: Spinnenseiden haben mechanische Eigenschaften, die jene der meisten natürlichen und synthetischen Fasern übertreffen. Wegen der Komplexität des In‐vivo‐Spinnprozesses scheiterte bislang seine Nachahmung durch klassische Spinnmethoden. Die Analyse des natürlichen Prozesses, kombiniert mit Erkenntnissen aus In‐vitro‐Untersuchungen, hilft bei der Entwicklung eines bionischen Spinnverfahrens zur Spinnenseidenherstellung (siehe Bild).magnified imageDie Proteinfaser Spinnenseide ist hinsichtlich ihrer Materialeigenschaften anderen natürlichen und vielen synthetischen Fasern überlegen. In der Natur beruht die Bildung einer festen Faser aus einem löslichen Spinnenseidenprotein auf komplexen biochemischen und physikalischen Prozessen, die in spezialisierten Spinnorganen ablaufen. In diesem Aufsatz wird der natürliche Prozess der Seidenherstellung ausgehend von der Gentranskription über die Proteinverarbeitung bis hin zur abschließenden Faserbildung vorgestellt; ebenso kommen technische Verfahren zur Seidenverspinnung zur Sprache. In‐vivo‐ und In‐vitro‐Befunde auf dem Gebiet der Spinnenseidenforschung bilden die Grundlage für die Gestaltung neuer Proteine und Verarbeitungsstrategien, die die Anwendung dieser faszinierenden proteinösen Materialien in der Technik und der Medizin ermöglichen können.

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Production and purification of recombinant DP1B silk-like protein in plants

Cited by 41 publications

References 18 publications

Recombinant Spider Silks—Biopolymers with Potential for Future Applications

Recombinant Spider Silks—Biopolymers with Potential for Future Applications

Biotechnological Production of Spider‐Silk Proteins Enables New Applications

Spinnenseide: vom löslichen Protein zur außergewöhnlichen Faser

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