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

3 Biotech
Tingting LiuFanghai Wang

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.

References

Jan 1, 1997·Applied Microbiology and Biotechnology·S R Fahnestock, S L Irwin
Jan 1, 1997·Applied Microbiology and Biotechnology·S R Fahnestock, L A Bedzyk
Jul 28, 2010·Proceedings of the National Academy of Sciences of the United States of America·Xiao-Xia XiaSang Yup Lee
Oct 15, 2013·Microbial Biotechnology·Olena TokarevaDavid L Kaplan

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