Lyons Russell E, Lesieur Emmanuelle, Kim Misook, Wong Darren C C, Huson Mickey G, Nairn Kate M, Brownlee Alan G, Pearson Roger D, Elvin Christopher M
CSIRO Livestock Industries, Queensland Bioscience Precinct, St Lucia, QLD 4072, Australia.
Protein Eng Des Sel. 2007 Jan;20(1):25-32. doi: 10.1093/protein/gzl050. Epub 2007 Jan 11.
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.
弹性蛋白是一种存在于昆虫表皮特定区域的弹性蛋白，具有独特的弹性和疲劳寿命特性。与许多弹性蛋白一样，包括弹性硬蛋白、麦醇溶蛋白和蜘蛛丝，弹性蛋白含有不同的重复结构域，这些结构域似乎赋予了蛋白质弹性特性。我们实验室最近的研究表明，克隆和表达果蝇CG15920基因的外显子1，该基因编码一种假定的类弹性蛋白，会产生一种重组蛋白，该蛋白可以通过光化学交联形成一种高弹性的弹性生物材料（Rec1弹性蛋白）。当前的研究描述了一种递归克隆策略，用于基于果蝇和冈比亚按蚊类弹性蛋白基因中的重复结构域，生成编码多个共有多肽拷贝的合成基因。还报道了一种可应用于这些合成蛋白和Rec1的简单非色谱纯化方法。这些设计和纯化类弹性蛋白周期性多肽的方法将有助于未来对弹性蛋白的结构和功能特性进行研究，以及开发新型高弹性生物材料。
