Purification and characterization of recombinant spider silk expressed in Escherichia coli.
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.
Arcidiacono
S
S
Biotechnology Division, U.S. Army Natick Army Research, Development, and Engineering Center, Natick, MA 01760, USA.
Mello
C
C
Kaplan
D
D
Cheley
S
S
Bayley
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics*; D019476:Insect Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D047011:Silk; D013112:Spiders
D018076:DNA, Complementary; D019476:Insect Proteins; D011994:Recombinant Proteins; D047011:Silk