The mechanical properties of chimeric silk are improved by expressing the full-length Trichonephila clavipes major ampullate spidroin gene in the silkworm Bombyx mori via recombinant AcMNPV

IF 3.3 2区 医学 Q2 ENGINEERING, BIOMEDICAL Journal of the Mechanical Behavior of Biomedical Materials Pub Date : 2024-09-12 DOI:10.1016/j.jmbbm.2024.106742
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Abstract

Spider silk is a type of natural protein fiber with excellent toughness and tensile strength. The mechanical properties of chimeric silk have been improved by integrating the spider silk protein gene into the silkworm (Bombyx mori) genome, but this strategy requires a long time to produce genetically modified silkworms. In this study, to rapidly produce chimeric silkworms/spider silk with improved toughness and tensile strength, recombinant Autographa californica multiple nucleopolyhedrovirus (AcMNPV), AcMNPV-FHP-MaSp-G, harboring a full-length Trichonephila clavipes major ampullate spidroin G (MaSp-G) gene driven by the silkworm fibroin heavy chain (Fib-H) promoter, was constructed, in which the signal peptide sequence of the MaSp-G gene was replaced by the signal peptide sequence of the Fib-H gene. Western blot and LC–MS/MS results showed that MaSp-G was successfully expressed in the posterior silk gland of silkworm larvae infected with AcMNPV-FHP-MaSp-G and secreted into the cocoon. Mechanical property tests revealed that the average maximum breaking stress and the average maximum elastic strain of chimeric silkworms/spider silk were 497.867 MPa and 14.824%, respectively, which were 36.53% and 23.55% greater than those of silk produced by normal silkworms. Fourier transform infrared (FTIR) spectroscopy revealed that the proportions of β-sheets, α-helices, and β-turns in the chimeric silk increased by 18.22%, 16.92%, and 18.72%, respectively. These results indicate that the mechanical properties of the chimeric silk produced by silkworms infected with AcMNPV-FHP-MaSp-G were significantly improved, which provides a new method for rapid production of chimeric silk in a genetically modified/genome-edited silkworm-independent manner.

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通过重组 AcMNPV 在家蚕中表达全长的克氏毛蛛主要安瓿鞘氨醇基因,改善嵌合蚕丝的机械性能
蛛丝是一种天然蛋白质纤维,具有极佳的韧性和抗拉强度。通过将蛛丝蛋白基因整合到家蚕(Bombyx mori)基因组中,可以改善嵌合蚕丝的机械性能,但这种策略需要很长时间才能培育出转基因家蚕。在本研究中,为了快速生产出具有更好韧性和抗拉强度的嵌合蚕/蜘蛛丝,重组了加州多核多角体病毒(AcMNPV),AcMNPV-FHP-MaSp-G、构建了由家蚕纤维蛋白重链(Fib-H)启动子驱动的重组家蚕多核多角体病毒(AcMNPV)--AcMNPV-FHP-MaSp-G,其中 MaSp-G 基因的信号肽序列被 Fib-H 基因的信号肽序列所取代。Western 印迹和 LC-MS/MS 结果表明,MaSp-G 在感染 AcMNPV-FHP-MaSp-G 的蚕幼虫后丝腺中成功表达并分泌到蚕茧中。力学性能测试表明,嵌合蚕/蜘蛛丝的平均最大断裂应力和平均最大弹性应变分别为 497.867 MPa 和 14.824%,比普通蚕丝分别高出 36.53% 和 23.55%。傅立叶变换红外光谱(FTIR)显示,嵌合蚕丝中β片、α螺旋和β匝的比例分别增加了18.22%、16.92%和18.72%。这些结果表明,感染 AcMNPV-FHP-MaSp-G 的家蚕生产的嵌合蚕丝的机械性能得到了显著改善,为不依赖转基因/基因组编辑的家蚕快速生产嵌合蚕丝提供了一种新方法。
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来源期刊
Journal of the Mechanical Behavior of Biomedical Materials
Journal of the Mechanical Behavior of Biomedical Materials 工程技术-材料科学:生物材料
CiteScore
7.20
自引率
7.70%
发文量
505
审稿时长
46 days
期刊介绍: The Journal of the Mechanical Behavior of Biomedical Materials is concerned with the mechanical deformation, damage and failure under applied forces, of biological material (at the tissue, cellular and molecular levels) and of biomaterials, i.e. those materials which are designed to mimic or replace biological materials. The primary focus of the journal is the synthesis of materials science, biology, and medical and dental science. Reports of fundamental scientific investigations are welcome, as are articles concerned with the practical application of materials in medical devices. Both experimental and theoretical work is of interest; theoretical papers will normally include comparison of predictions with experimental data, though we recognize that this may not always be appropriate. The journal also publishes technical notes concerned with emerging experimental or theoretical techniques, letters to the editor and, by invitation, review articles and papers describing existing techniques for the benefit of an interdisciplinary readership.
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