Laboratory of Molecular Microbiology and Virology, Department of Health Sciences, Faculty of Medicine and Surgery, University of Milano, Bicocca, Via Cadore, 48, 20900 Monza, MB, Italy.
Mol Biotechnol. 2013 Jul;54(3):818-28. doi: 10.1007/s12033-012-9629-1.
The purpose of this study was the output and set up of the milk array, a dedicated array designed to investigate the expression levels of many genes involved in cow mammary gland inflammation and milk production regulation. First, a new targeted genes panel was selected. Successively, the microarray reliability was examined by yellow and dye swap experiments using the normal and mastitic mammary gland samples from the same cow. The sensitivity and reliability were evaluated using different amounts of the same mastitic mammary gland RNA: a good linear regression (R (2) = 0.758) was obtained also using only 3 μg of RNA. We used both reverse transcriptase RT-qPCR and the microarray to analyze 100 bovine genes (96 known to be involved in inflammation and milk production regulation and four housekeeping genes) in pooled total RNA isolated from tissue samples. All genes were detectable by RT-qPCR and microarray: a good mean correlation coefficient over all samples of 0.885 showed that both methods were similarly well suited to analyze gene expression in these samples. This report describes the development of small DNA microarray of fully defined genes suitable for analysis of expression of many genes involved in cow mammary gland inflammation and milk production regulation; this platform will prove useful as diagnostic tool prototype to perform a more in-depth analysis of the milk quality and mammary glands health status.
本研究的目的是输出和建立牛奶阵列，这是一种专门设计的阵列，用于研究涉及奶牛乳腺炎症和乳汁分泌调节的许多基因的表达水平。首先，选择了一个新的靶向基因面板。然后，使用来自同一头奶牛的正常和乳腺炎乳腺样本进行黄色和染料交换实验来检查微阵列的可靠性。使用不同量的相同乳腺炎乳腺 RNA 评估了灵敏度和可靠性：即使使用仅 3μg 的 RNA，也可以获得良好的线性回归（R (2) = 0.758）。我们使用逆转录 RT-qPCR 和微阵列分析了从组织样本中分离的总 RNA 中 100 个牛基因（96 个已知参与炎症和乳汁分泌调节，还有 4 个管家基因）。所有基因都可以通过 RT-qPCR 和微阵列检测到：所有样品的平均相关系数为 0.885，表明两种方法都非常适合分析这些样品中的基因表达。本报告描述了完全定义基因的小型 DNA 微阵列的开发，该微阵列适用于分析涉及奶牛乳腺炎症和乳汁分泌调节的许多基因的表达；该平台将作为诊断工具原型，用于更深入地分析牛奶质量和乳腺健康状况。
