Liu Frank Y C, Liu Jessica Y X, Yao Xiu, Wang Boxiang
Department of Biology, Link-Spider Co. Ltd., Room D-E, Floor 22, Caifu Building, Fuhua 3rd Rd., Shenzhen, Guangdong 518000, China; Science Department, Newton South High School, 140 Brandeis Rd., Newton, MA 02459, USA.
Department of Biology, Link-Spider Co. Ltd., Room D-E, Floor 22, Caifu Building, Fuhua 3rd Rd., Shenzhen, Guangdong 518000, China; Department of Material Science and Engineering, College of Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, PA 15213, USA.
Int J Biol Macromol. 2022 Mar 1;200:362-369. doi: 10.1016/j.ijbiomac.2021.12.078. Epub 2021 Dec 30.
Araneid spider silk glands can spin seven silk types that have task-specific properties owing to the higher order structure of spider silk proteins. This gives silks superior potential as novel biomaterials. Nephila pilipes, the giant golden orb-weaver, is one of the largest spiders and spins silk with exceptional torsional deformation, toughness, and other properties to support its mass; further investigation relies on a complete amino acid sequence. However, there are no full-length N. pilipes spidroin sequences; in fact, across species, most sequences remain fragmentary because of repetitive region assembly difficulties in short-read sequencing. Here, we develop a hybrid sequencing method that utilizes short-read sequencing to identify seven spidroin terminals in N. pilipes, and long-read sequencing to confirm the full-length pyriform spidroin 1 (PySp1) gene. PySp1 is 11,181 base pairs, with a single exon encoding a 3,726 amino acid protein, the QQ(x)Qx motif, and lower repeat homogenization, distinct characteristics of genera Nephilinae PySp1. The full-length N. pilipes PySp1 sequences sheds light on spidroin evolution and demonstrates a helpful strategy to find full-length spidroins.
园蛛科蜘蛛的丝腺能够纺出七种具有特定任务属性的丝，这归因于蜘蛛丝蛋白的高级结构。这使得蜘蛛丝作为新型生物材料具有卓越的潜力。大木林蛛是最大的蜘蛛之一，它所纺出的丝具有非凡的抗扭变形能力、韧性及其他特性以支撑其体重；进一步的研究依赖于完整的氨基酸序列。然而，目前尚无大木林蛛蛛丝蛋白的全长序列；事实上，在所有物种中，由于短读长测序中重复区域组装困难，大多数序列仍然是片段化的。在此，我们开发了一种混合测序方法，利用短读长测序来识别大木林蛛的七种蛛丝蛋白末端，并利用长读长测序来确认全长梨状蛛丝蛋白1（PySp1）基因。PySp1基因全长11,181个碱基对，有一个外显子编码一个含有3,726个氨基酸的蛋白质、QQ(x)Qx基序，且重复序列同质化程度较低，这些是络新妇亚科PySp1的独特特征。大木林蛛PySp1的全长序列为蛛丝蛋白的进化提供了线索，并展示了一种寻找全长蛛丝蛋白的有效策略。
