Marques M Carmen, Perez-Amador Miguel A
Instituto de Biología Molecular y Celular de Plantas-IBMCP, Universidad Politécnica de Valencia-UPV and Consejo Superior de Investigaciones Científicas-CSIC, CPI 8E, Ingeniero Fausto Elio s/n, Valencia 46022, Spain.
Methods Mol Biol. 2012;815:51-65. doi: 10.1007/978-1-61779-424-7_5.
We have developed an integrated method to generate a normalized cDNA collection enriched in full-length and rare transcripts from citrus, using different species and multiple tissues and developmental stages. Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. In this regard, the availability of full-length cDNA clones facilitates functional analysis of the corresponding genes enabling manipulation of their expression and the generation of a variety of tagged versions of the native protein. The development of full-length cDNA sequences has the power to improve the quality of genome annotation, as well as provide tools for functional characterization of genes.
我们开发了一种综合方法，利用不同的物种、多个组织和发育阶段，从柑橘中生成富含全长和稀有转录本的标准化cDNA文库。解读现代基因组测序技术产生的不断增加的原始序列信息面临多重挑战，如基因功能分析和基因组注释。在这方面，全长cDNA克隆的可用性有助于对相应基因进行功能分析，从而能够操纵其表达并生成多种天然蛋白质的标记版本。全长cDNA序列的开发有助于提高基因组注释的质量，并为基因的功能表征提供工具。
