Name: synthetic spider silk production on a laboratory scale
Uploaded: 2012-07-18T12:00:00
Description: Watch this Scientific Journal Video about Synthetic Spider Silk Production on a Laboratory Scale at JoVE.com

这项工作由芮NSF的资助小型断路器-0950372和DMR-1105310题为“黑寡妇蜘蛛丝和力学性能蜘蛛胶彩衣的分子鉴定”，分别支持。

<img alt="图11" src="/files/ftp_upload/4191/4191fig1overview.jpg" /> 图形概述：纺纱过程的仿生学 仿生学的天然蜘蛛丝的生产途径：制造人造丝的路线此图像显示了主壶腹腺金球织布， 蜘蛛clavipes，天然蚕丝生产（白色文字）利用组件。尾地区综合大量的丝蛋白，被运送到壶腹，为纺丝原液的存储区域。这种集中的涂料是通过纺纱管解决方案体验离子交换和?...

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这种方法纺成的合成纤维是机械上的同一量级的天然纤维相比。通过假脱机和自旋后抽签过程机械化，减少人为错误的数量，实验样品之间的变化有更多的控制，大大降低了。 我们的方法提供了潜力调查其他纺蜘蛛基因家族的其他成员的cDNA编码的重组蛋白纤维的力学性能。潜在的，它可以决定不同的蛋白质模块内素，或不同的素类型之间的机械作用，23它还允许蚕丝...

蜘蛛丝具有非凡的机械性能，包括高强度钢，芳纶和尼龙出执行数的人造材料，旋转1蜘蛛至少6-7种不同的光纤类型，显示不同的力学性能，数额不等的拉伸强度和可扩展性设计执行特定的生物任务。2研究的科学家正在迅速推行的蜘蛛丝作为下一代生物材料由于其优异的力学性能，其生物相容性，无毒，绿色材料的性质使用。由于“吃人的3,4和有毒的蜘蛛纲的性质，通过耕种收割蜘蛛丝是不是一个实际的战略，以满足工业规模生产所必需的要求。因此，科学家们转向了从合成纤维纺丝加上在体外重组丝蛋白的转基因生物的生产5-8全长重组蜘蛛丝蛋白表达本身的纯化蛋白质。一直技术上的困难，由于他们的基因序列的内在属性，其中包括他们的高度重复的性质和物理长度（15 KB），GC丰富的内容和偏见丙氨酸和甘氨酸密码子使用9-11迄今为止，大多数实验室都集中表达形式截断主壶腹丝蛋白的部分cDNA序列或人工合成的基因MaSp1或MaSp2 12-15纺纱合成蜘蛛丝是一个具有挑战性的过程，需要掌握知识，在若干学科，纺纱过程的复杂性没有得到充分暴露广大市民通过视频表示。事实上，只有少数遍布全球的实验室具备专业知识，表达蜘蛛丝的cDNA，净化丝蛋白，纺合成纤维和执行后的自旋战平，然后最后测试其生物材料的性能8。16,17纺合成纤维不同的方法包括湿法和干法纺丝以及静电方法16,18,19的所有程序，有一个共同的目标-一个协议，生产合成蜘蛛丝力学性能发展，对手自然线程大规模商业化生产进程。 在这里，我们描述的过程来生成实验室规模的人工蜘蛛丝，用湿纺方法。相对于其他纺纱方法，湿法纺丝纤维分析的最一致的结果。我们概括与表达中的细菌其纯化重组丝蛋白，其次这个过程的开始，然后纺纱，包括自旋后抽签的方法适用于“纺”的纤维，产生线程的描述蛋白质的准备步骤材料性能接近天然蜘蛛丝的质量。我们的方法论的y的设计紧密地模仿天然蚕丝纤维的纺纱工艺和倚重它吸引我们的ORB和芯织网的蜘蛛。20-22此外，丝绸生产的腺体结构和功能方面的专长，我们的结论与必要的步骤，以确定使用tensometer绘制应力 - 应变曲线，这让研究者计算的极限强度，极限应变，韧性纤维，合成纤维材料的性能。最后，但显著的价值，纺纱，后台处理和绘图仪器可以在家使用市售的部分，而不是购买定制的精致和昂贵的设备，建造。

<table border="1"><tbody><tr><td> 试剂/设备 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> PBAD / TOPO ThioFusion的表达试剂盒</td><td> Invitrogen公司</td><td> K370-01 </td><td></td></tr><tr><td>的快攻细胞裂解试剂，10X </td><td> Promega公司</td><td> V857C </td><td></td></tr><tr><td>镍-NTA琼脂糖</td><td> Qiagen公司</td><td> 30210 </td><td>包括对缓冲区的说明</td></tr><tr><td> ProteoSilver银染色套件</td><td> Sigma-Aldrich公司</td><td> PROTSIL1-1KT </td><td></td></tr><tr><td>自由贸易区冻干机</td><td> labconco </td><td> 7960041 </td><td>自由贸易区12Plus </td></tr><tr><td>六氟异丙醇（异丙醇） </td><td> Sigma-Aldrich公司</td><td> 52512 </td><td></td></tr><tr><td>注射器</TD&gt; <td>汉密尔顿</td><td> 7657-01 </td><td> 250微升</td></tr><tr><td>针</td><td>汉密尔顿</td><td> 7780-01 </td><td> 26S计，钝端可拆卸针</td></tr><tr><td>注射泵</td><td>哈佛仪器</td><td> 702208 </td><td> 11Plus </td></tr><tr><td>数显卡尺</td><td>卡雷拉</td><td> CP5906 </td><td> 0-150毫米范围</td></tr><tr><td>不锈钢钢钳</td><td>世界精密仪器</td><td> 501764 </td><td>迷你杜蒙＃M5S </td></tr><tr><td>马达</td><td>自然轧机</td><td> 7090529 </td><td> 12VDC，2 rpm的转速</td></tr><tr><td>线性驱动器</td><td>华纳电</td><td> 01-D024-0050-A06-LP的，符合IP65 </td><td> 24VDC，6英寸的范围</td></tr><tr><td>解剖显微镜</td><td>徕卡公司</td><td>徕卡MZ16 </td><td></td></tr><tr><td>数字microscOPE相机</td><td>徕卡公司</td><td> DFC320 </td><td>软件：徕卡应用套件v2.8.1 </td></tr><tr><td> vannas剪刀</td><td>世界精密仪器</td><td> 500260 </td><td></td></tr><tr><td> microtensometer </td><td>极光科学</td><td> 310C </td><td> ：5N双模式系统</td></tr></tbody></table>

synthetic spider silk production on a laboratory scale

随着社会的进步和资源日益稀少，它正变得越来越重要，以培育新技术，新一代生物材料具有较高的性能工程师。这些新的结构材料的发展必须是快速，具有成本效益和涉及的加工方法和产品，环保和可持续发展。蜘蛛能纺出多种不同类型的纤维，具有不同的力学性能，提供丰富的新一代工程材料的来源，仿生学，对手最好的人造和天然材料。由于收集大量天然蜘蛛丝是不切实际的，人造丝生产有能力提供无限量供应的线程访问科学家。因此，如果纺纱过程可以简化和完善，人工蜘蛛纤维的防弹衣，应用范围广泛，手术缝合线的潜在用途，绳索和电缆，轮胎，乐器的弦，和航空航天技术的复合材料。以推动人造丝生产过程中产生纤维，显示在他们的物质属性从低自旋旋转方差，我们开发湿纺协议，整合重组蜘蛛丝蛋白的蛋白质的细菌，净化和浓度的表达其次是纤维挤压和机械旋转后处理。这是第一个可视化表示形式，揭示了一步一步的过程，旋转和分析实验室规模的人造丝纤维。它还提供了详细资料，以尽量减少从同一纺丝液纺纤维中引入变异。总的来说，这些方法将推动人造丝生产过程中，导致更高的质量超过天然蜘蛛丝纤维。

Watch this Scientific Journal Video about Synthetic Spider Silk Production on a Laboratory Scale at JoVE.com

Synthetic Spider Silk Production on a Laboratory Scale

尽管优秀的机械和生化蜘蛛丝的性能，这种材料可以收获大量的常规手段。在这里，我们描述了一个有效的战略，以旋转人工蜘蛛丝纤维，这是一个重要研究蜘蛛丝作为下一代生物材料的生产和使用的调查过程。

实验室规模的人工合成蜘蛛丝生产

As society progresses and resources become scarcer, it is becoming increasingly important to cultivate new technologies that engineer next generation ...

Research

JoVE Journal

Bioengineering

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

高通量微RNA分析：优化复纳升秤定量RT - PCR Fluidig​​m公司动态ArrayTM国际金融中心

The broad involvement of miRNAs in critical processes underlying development, tissue homoeostasis and disease has led to a surging interest among the ...

科研

生物工程

Engineering Adherent Bacteria by Creating a Single Synthetic Curli Operon

通过创建一个的合成Curli的操纵子工程粘附的细菌

The  method described here consists in redesigning E. coli adherence properties by assembling the minimum number of  curli genes under the control of a ...

Air Filter Devices Including Nonwoven Meshes of Electrospun Recombinant Spider Silk Proteins

空气过滤设备，包括非织造网格静电重组蜘蛛丝蛋白

Based on the natural sequence of Araneus diadematus Fibroin 4 (ADF4), the recombinant spider silk protein eADF4(C16) has been engineered. This highly ...

Designing Silk-silk Protein Alloy Materials for Biomedical Applications

设计丝绸蚕丝蛋白合金生物医用材料

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, ...

Production and Targeting of Monovalent Quantum Dots

生产和目标单价量子点

The multivalent nature of commercial quantum dots (QDs) and the difficulties associated with producing monovalent dots have limited their applications in ...

Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5

基于人类腺病毒5型高容量腺病毒载体的克隆及规模化生产

High-capacity adenoviral vectors (HCAdV) devoid of all viral coding sequences represent one of the most advanced gene delivery vectors due to their high ...

Engineered 3D Silk-collagen-based Model of Polarized Neural Tissue

极化神经组织工程三维蚕丝胶原蛋白为基础的模式

Despite huge efforts to decipher the anatomy, composition and function of the brain, it remains the least understood organ of the human body. To gain a ...

Manufacture and Drug Delivery Applications of Silk Nanoparticles

纳米丝绸制造和药物输送应用

Silk is a promising biopolymer for biomedical and pharmaceutical applications due to its outstanding mechanical properties, biocompatibility and ...

Power Input Measurements in Stirred Bioreactors at Laboratory Scale

实验室规模搅拌生物反应器的功率输入测量

The power input in stirred bioreactors is an important scaling-up parameter and can be measured through the torque that acts on the impeller shaft during ...

Material Formation of Recombinant Spider Silks through Aqueous Solvation using Heat and Pressure

热、压水固化重组蜘蛛丝的材料形成

Many spiders produce seven types of silks. Six of the silks are fiber in form when produced by the spiders. These fibers are not water soluble. In order ...