The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
Construction, Cloning, and Expression of Synthetic Genes Encoding Spider Dragline Silk
Abstract
Synthetic genes encoding recombinant spider silk proteins have been constructed, cloned, and expressed. Protein sequences were derived from Nephila clavipes dragline silk proteins and reverse-translated to the corresponding DNA sequences. Codon selection was chosen to maximize expression levels in Escherichia coli. DNA "monomer" sequences were multimerized to encode high molecular weight synthetic spider silks using a "head-to-tail" construction strategy. Multimers were cloned into a prokaryotic expression vector and the encoded silk proteins were expressed in E. coli upon induction with IPTG. Four multimer, ranging in size from 14.7 to 41.3 kDa, were chosen for detailed analysis. These proteins were isolated by immobilized metal affinity chromatography and purified using reverse-phase HPLC. The composition and identity of the purified proteins were confirmed by amino acid composition analysis, N-terminal sequencing, laser desorption mass spectroscopy, and Western analysis using antibodies reactive to native spider dragline silk. Circular dichroism measurements indicate that the synthetic spider silks have substantial beta-sheet structure.
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Citations
Cites
A rapid alkaline extraction procedure for screening recombinant plasmid DNA
Birnboim, H. C., & Doly, J. (1979) · Nucleic Acids Research · doi:10.1093/nar/7.6.1513 · Cited by 14,686
Cited by
Altman, G. H., Diaz, F., Jakuba, C., Calabro, T., Horan, R. L., Chen, J., Lu, H., Richmond, J., & Kaplan, D. L. (2003) · Biomaterials · doi:10.1016/s0142-9612(02)00353-8 · Cited by 3,458
Self-citationLiquid crystalline spinning of spider silk
Vollrath, F., & Knight, D. P. (2001) · Nature · doi:10.1038/35069000 · Cited by 1,610
New Opportunities for an Ancient Material
Omenetto, F. G., & Kaplan, D. L. (2010) · Science · doi:10.1126/science.1188936 · Cited by 1,457
Self-citationFunctionalized silk-based biomaterials for bone formation
Sofia, S., McCarthy, M. B., Gronowicz, G., & Kaplan, D. L. (2001) · Journal of Biomedical Materials Research · doi:10.1002/1097-4636(200101)54:1<139::aid-jbm17>3.0.co;2-7 · Cited by 819
Self-citationThe Biomedical Use of Silk: Past, Present, Future
Holland, C., Numata, K., Rnjak‐Kovacina, J., & Seib, F. P. (2019) · Advanced Healthcare Materials · doi:10.1002/adhm.201800465 · Cited by 815
Metadata
- Authors
- Prince, J. T., McGrath, K. P., DiGirolamo, C. M., & Kaplan, D. L.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1995-08-29
- Volume
- 34
- Issue
- 34
- Pages
- 10879-10885
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2085830562
- Cited by
- 282
- References
- 1
- FWCI
- 7.07
- Type
- journal article
- Language
- en
- Open access
- closed
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- Prince, J. T., McGrath, K. P., DiGirolamo, C. M., & Kaplan, D. L. (1995). Construction, Cloning, and Expression of Synthetic Genes Encoding Spider Dragline Silk. Biochemistry. https://doi.org/10.1021/bi00034a022
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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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