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Construction of a synthetic Araneus ventricosus dragline silk gene multimer and its expression in Escherichia coli

Authors
  • Liu, Tingting1
  • Liang, Anwen1
  • Liang, Ziqiang1
  • Li, Guanghong1
  • Wang, Fanghai1
  • 1 Sun Yat-sen University, State Key Laboratory for Biocontrol and Institute of Entomology, Guangzhou, 510275, China , Guangzhou (China)
Type
Published Article
Journal
3 Biotech
Publisher
Springer International Publishing
Publication Date
May 11, 2018
Volume
8
Issue
5
Identifiers
DOI: 10.1007/s13205-018-1285-0
Source
Springer Nature
Keywords
License
Yellow

Abstract

One of the most representative core gene sequence of Araneus ventricosus dragline silk protein partial cDNA monomer (JN857964.2) was selected and multimerized using a “head-to-tail” strategy by compatible but nonregenerable sites at both ends resulting in a concatemer of 16 contiguous monomers. This concatemer was cloned into pET-28a(+) expression vector and transformed into Escherichia coli. A 52.6 kDa silk protein was successfully expressed and detected by SDS–PAGE and confirmed by Western blotting. A maximum yield of the silk protein was expressed with 7.06 mM IPTG after 5 h incubation. This is the first report on the construction and overexpression of a A. ventricosus dragline silk multimeric gene construct and the results from our study will provide a reference point for further exploration and development of large-scale production of spider silk protein.

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