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生物黏附研究中的分子生物学方法。

Molecular biology approaches in bioadhesion research.

机构信息

University of Innsbruck, Institute of Zoology and Center for Molecular Biosciences Innsbruck, Technikerstraße 25, A-6020 Innsbruck, Austria.

出版信息

Beilstein J Nanotechnol. 2014 Jul 8;5:983-93. doi: 10.3762/bjnano.5.112. eCollection 2014.

Abstract

The use of molecular biology tools in the field of bioadhesion is still in its infancy. For new research groups who are considering taking a molecular approach, the techniques presented here are essential to unravelling the sequence of a gene, its expression and its biological function. Here we provide an outline for addressing adhesion-related genes in diverse organisms. We show how to gradually narrow down the number of candidate transcripts that are involved in adhesion by (1) generating a transcriptome and a differentially expressed cDNA list enriched for adhesion-related transcripts, (2) setting up a BLAST search facility, (3) perform an in situ hybridization screen, and (4) functional analyses of selected genes by using RNA interference knock-down. Furthermore, latest developments in genome-editing are presented as new tools to study gene function. By using this iterative multi-technologies approach, the identification, isolation, expression and function of adhesion-related genes can be studied in most organisms. These tools will improve our understanding of the diversity of molecules used for adhesion in different organisms and these findings will help to develop innovative bio-inspired adhesives.

摘要

在生物黏附领域应用分子生物学工具仍处于起步阶段。对于那些考虑采用分子方法的新研究小组来说,这里介绍的技术对于揭示基因序列、其表达和生物学功能是必不可少的。在这里,我们提供了一个针对不同生物体内与黏附相关基因的研究大纲。我们展示了如何通过以下步骤逐渐减少涉及黏附的候选转录本的数量:(1)生成转录组和富含黏附相关转录本的差异表达 cDNA 列表,(2)建立 BLAST 搜索工具,(3)进行原位杂交筛选,以及(4)通过 RNA 干扰敲低对选定基因进行功能分析。此外,还介绍了基因组编辑的最新进展,作为研究基因功能的新工具。通过使用这种迭代的多技术方法,可以在大多数生物体中研究与黏附相关的基因的鉴定、分离、表达和功能。这些工具将增进我们对不同生物体中用于黏附的分子多样性的理解,这些发现将有助于开发创新的仿生黏附剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/94f3/4142862/4096ec9389ce/Beilstein_J_Nanotechnol-05-983-g002.jpg

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