Design and facile production of recombinant resilin-like polypeptides: gene construction and a rapid protein purification method.
Resilin is an elastic protein found in specialized regions of the cuticle of insects, which displays unique resilience and fatigue lifetime properties. As is the case with many elastomeric proteins, including elastin, gliadin and spider silks, resilin contains distinct repetitive domains that appear to confer elastic properties to the protein. Recent work within our laboratory has demonstrated that cloning and expression of exon 1 of the Drosophila melanogaster CG15920 gene, encoding a putative resilin-like protein, results in a recombinant protein that can be photochemically crosslinked to form a highly resilient, elastic biomaterial (Rec1 resilin). The current study describes a recursive cloning strategy for generating synthetic genes encoding multiple copies of consensus polypeptides, based on the repetitive domains within resilin-like genes from D. melanogaster and Anopheles gambiae. A simple non-chromatographic purification method that can be applied to these synthetic proteins and Rec1 is also reported. These methods for the design and purification of resilin-like periodic polypeptides will facilitate the future investigation of structural and functional properties of resilin, and the development of novel highly resilient biomaterials.
Lyons
Russell E
RE
CSIRO Livestock Industries, Queensland Bioscience Precinct, St Lucia, QLD 4072, Australia. Russell.Lyons@csiro.au
Lesieur
Emmanuelle
E
Kim
Misook
M
Wong
Darren C C
DC
Huson
Mickey G
MG
Nairn
Kate M
KM
Brownlee
Alan G
AG
Pearson
Roger D
RD
Elvin
Christopher M
CM
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000235:genetics; D001483:Base Sequence; D004331:Drosophila melanogaster / Q000235:genetics; D004548:Elasticity; D020724:Elastomers; D004591:Electrophoresis, Polyacrylamide Gel; D019476:Insect Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D020724:Elastomers; D019476:Insect Proteins; D011994:Recombinant Proteins; C030951:resilin