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激光辅助显微切割技术,一种用于了解植物组织特异性基因表达模式和功能基因组学的有效技术。

Laser assisted microdissection, an efficient technique to understand tissue specific gene expression patterns and functional genomics in plants.

作者信息

Gautam Vibhav, Sarkar Ananda K

机构信息

National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi, 110067, India.

出版信息

Mol Biotechnol. 2015 Apr;57(4):299-308. doi: 10.1007/s12033-014-9824-3.

Abstract

Laser assisted microdissection (LAM) is an advanced technology used to perform tissue or cell-specific expression profiling of genes and proteins, owing to its ability to isolate the desired tissue or cell type from a heterogeneous population. Due to the specificity and high efficiency acquired during its pioneering use in medical science, the LAM technique has quickly been adopted for use in many biological researches. Today, it has become a potent tool to address a wide range of questions in diverse field of plant biology. Beginning with comparative transcriptome analysis of different tissues such as reproductive parts, meristems, lateral organs, roots etc., LAM has also been extensively used in plant-pathogen interaction studies, proteomics, and metabolomics. In combination with next generation sequencing and proteomics analysis, LAM has opened up promising opportunities in the area of large scale functional studies in plants. Ever since the advent of this technique, significant improvements have been achieved in term of its instrumentation and method, which has made LAM a more efficient tool applicable in wider research areas. Here, we discuss the advancement of LAM technique with special emphasis on its methodology and highlight its scope in modern research areas of plant biology. Although we put emphasis on use of LAM in transcriptome studies, which is mostly used, we also discuss its recent application and scope in proteome and metabolome studies.

摘要

激光辅助显微切割(LAM)是一项先进技术,由于其能够从异质群体中分离出所需的组织或细胞类型,因而可用于对基因和蛋白质进行组织或细胞特异性表达谱分析。由于在医学科学中的开创性应用所获得的特异性和高效率,LAM技术已迅速被应用于许多生物学研究中。如今,它已成为解决植物生物学不同领域中广泛问题的有力工具。从对不同组织(如生殖器官、分生组织、侧生器官、根等)的比较转录组分析开始,LAM也已广泛应用于植物-病原体相互作用研究、蛋白质组学和代谢组学。与下一代测序和蛋白质组学分析相结合,LAM在植物大规模功能研究领域开辟了广阔前景。自该技术问世以来,其仪器设备和方法已取得了显著改进,这使得LAM成为一种更高效的工具,可应用于更广泛的研究领域。在此,我们讨论LAM技术的进展,特别强调其方法,并突出其在植物生物学现代研究领域中的应用范围。尽管我们重点讨论LAM在转录组研究中的应用(这是其最常用的用途),但我们也讨论了它在蛋白质组和代谢组研究中的最新应用及范围。

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