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

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.

SUBMITTER: Liu T

PROVIDER: S-EPMC5948186 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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

Liu Tingting T Liang Anwen A Liang Ziqiang Z Li Guanghong G Wang Fanghai F

3 Biotech 20180511 5

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 ...[more]

PMID: 29765810

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