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土耳其骨骼肌 6K 寡核苷酸微阵列的特征描述。

Characterization of a 6K oligonucleotide turkey skeletal muscle microarray.

机构信息

Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48824, USA.

出版信息

Anim Genet. 2011 Feb;42(1):75-82. doi: 10.1111/j.1365-2052.2010.02085.x.

Abstract

Consumer demand for lean, inexpensive meat products has driven the domestic turkey (Meleagris gallopavo) industry to unprecedented production; however, this has coincided with an increase in growth-induced myopathies and meat quality defects. With the aim of developing a new tool for the study of turkey growth and development at the muscle transcriptome level, a 6K oligonucleotide microarray was constructed, the Turkey Skeletal Muscle Long Oligo (TSKMLO) microarray. Skeletal muscle samples were collected at three critical stages in muscle development: 18-day embryo (hyperplasia), 1-day post-hatch (hypertrophy), and 16-week (market age) from two genetic lines of turkeys: RBC2, a line maintained without selection pressure, and F, a line selected from the RBC2 line for increased 16-week body weight. Oligonucleotides were designed from sequences obtained from skeletal muscle cDNA libraries from the three developmental stages. Several unique controls, including mismatch and distance controls and scrambled sequences, were designed for 30 genes. Quality control hybridizations were completed, confirming the validity and repeatability of the array. Control features were evaluated across two larger experiments comparing developmental stage within genetic line or genetic line within each developmental stage, totaling 70 arrays. Mismatch and scrambled sequences appeared to be useful controls of specific hybridization for most genes. In addition, quantitative real-time RT-PCR confirmed microarray results. This creation and assessment of the TSKMLO array provides a valuable community resource for the study of gene expression changes related to turkey muscle growth and development.

摘要

消费者对低脂、廉价肉类产品的需求推动了国内火鸡(Meleagris gallopavo)产业达到了前所未有的生产水平;然而,这与生长诱导的肌肉病和肉质缺陷的增加同时发生。为了开发一种新的工具来研究火鸡在肌肉转录组水平上的生长和发育,构建了一个 6K 寡核苷酸微阵列,即 Turkey Skeletal Muscle Long Oligo(TSKMLO)微阵列。从两个火鸡遗传品系:RBC2(未受选择压力维持的品系)和 F(从 RBC2 品系中选择的 16 周体重增加的品系)中收集了三个肌肉发育关键阶段的骨骼肌样本:18 日龄胚胎(增生)、1 日龄出雏(肥大)和 16 周龄(市场年龄)。寡核苷酸是从三个发育阶段的骨骼肌 cDNA 文库中获得的序列设计的。为 30 个基因设计了几个独特的对照,包括错配和距离对照以及乱序序列。完成了质量控制杂交,证实了阵列的有效性和可重复性。在比较遗传品系内的发育阶段或遗传品系内的每个发育阶段的两个更大的实验中评估了对照特征,总共有 70 个阵列。错配和乱序序列似乎是大多数基因特定杂交的有用对照。此外,定量实时 RT-PCR 证实了微阵列结果。TSKMLO 阵列的创建和评估为研究与火鸡肌肉生长和发育相关的基因表达变化提供了有价值的社区资源。

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