ORIGINAL PAPER

Applied Microbiology and Biotechnology

, Volume 49, Issue 1, pp 31-38

First online:

Purification and characterization of recombinant spider silk expressed in Escherichia coli

  • S. ArcidiaconoAffiliated withBiotechnology Division, U.S. Army Natick Army Research, Development, and Engineering Center, Natick, MA 01760, USA Tel.: +1 (508) 233 5513; Fax: +1 (508) 233 5521
  • , C. MelloAffiliated withBiotechnology Division, U.S. Army Natick Army Research, Development, and Engineering Center, Natick, MA 01760, USA Tel.: +1 (508) 233 5513; Fax: +1 (508) 233 5521
  • , D. KaplanAffiliated withTufts University, Medford, MA 02155, USA
  • , S. CheleyAffiliated withTexas A&M University, College station, TX 77843, USA
  • , H. BayleyAffiliated withTexas A&M University, College station, TX 77843, USA

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

Abstract

A partial cDNA clone, from the 3′ end of the dragline silk gene was isolated from Nephila clavipes major ampullate glands. This clone contains a 1.7-kb insert, consisting of a repetitive coding region of 1.4-kb and a 0.3-kb nonrepetitive coding region; 1.5-kb of the 1.7-kb fragment was cloned into Escherichia coli and a␣43-kDa recombinant silk protein was expressed. Characterization of the purified protein by Western blot, amino acid composition analysis, and matrix-assisted laser desorption ionization/time-of-flight mass spectrometry confirms it to be spider dragline silk.