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用于反刍动物繁殖和生产(牛奶和牛肉)性状相关组织中基因表达谱分析的cDNA宏阵列资源。

A cDNA macroarray resource for gene expression profiling in ruminant tissues involved in reproduction and production (milk and beef) traits.

作者信息

Bernard C, Degrelle S, Ollier S, Campion E, Cassar-Malek I, Charpigny G, Dhorne-Pollet S, Hue I, Hocquette J-F, Le Provost F, Leroux C, Piumi F, Rolland G, Uzbekova S, Zalachas E, Martin P

机构信息

INRA, Unité de Recherche sur les Herbivores, Theix, 63122 Saint-Genès Champanelle, France.

出版信息

J Physiol Pharmacol. 2005 Jun;56 Suppl 3:215-24.

Abstract

cDNA arrays have proven to be useful tools to screen gene expression in many animal species including livestock species. A collaborative program was launched to construct a ruminant cDNA collection, representative of three tissues: Muscle, Embryo and Mammary gland, named MEM. This collection gathers clones mainly arising from 3 non-normalised cDNA libraries: a directed bovine muscle library, a 14-day-old bovine embryo library and a goat lactating mammary library. It is made up of 1896 clones (637 muscle, 882 embryo and 377 mammary cDNAs), selected after sequencing and bioinformatic analyses. Amplification products yielded from these clones as well as controls were printed onto Nylon membranes to generate macroarrays. Hybridisation with relevant cDNA targets allowed checking the location of about 50 cDNAs and the specificity of each sub-set of the repertoire. Macroarrays were hybridised with radiolabelled cDNA complex targets from five different tissues (muscle, embryo, mammary gland, adipose tissue and oocyte). Both somatic and germinal complex targets gave valid hybridisation signals with 45 to 80% of the printed probes. This specific cDNA collection now provides a powerful tool for transcriptomic studies with the ultimate objective to better understand physiological and metabolic functions in ruminants. It will be subsequently included into a forthcoming larger collection.

摘要

cDNA阵列已被证明是筛选包括家畜物种在内的许多动物物种基因表达的有用工具。启动了一个合作项目来构建一个反刍动物cDNA文库,该文库代表肌肉、胚胎和乳腺这三种组织,名为MEM文库。该文库收集的克隆主要来自3个非标准化cDNA文库:一个定向牛肌肉文库、一个14日龄牛胚胎文库和一个山羊泌乳期乳腺文库。它由1896个克隆组成(637个肌肉cDNA、882个胚胎cDNA和377个乳腺cDNA),这些克隆是经过测序和生物信息学分析后挑选出来的。从这些克隆以及对照中产生的扩增产物被打印到尼龙膜上以生成宏阵列。与相关cDNA靶标的杂交可以检查约50个cDNA的位置以及文库中每个子集的特异性。宏阵列与来自五个不同组织(肌肉、胚胎、乳腺、脂肪组织和卵母细胞)的放射性标记cDNA复合靶标进行杂交。体细胞和生殖细胞复合靶标与45%至80%的打印探针都产生了有效的杂交信号。这个特定的cDNA文库现在为转录组学研究提供了一个强大的工具,其最终目标是更好地了解反刍动物的生理和代谢功能。随后它将被纳入即将推出的更大的文库中。

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