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牛巨噬细胞特异性cDNA微阵列的开发与验证

Development and validation of a bovine macrophage specific cDNA microarray.

作者信息

Jensen Kirsty, Talbot Richard, Paxton Edith, Waddington David, Glass Elizabeth J

机构信息

Division of Genetics & Genomics, Roslin Institute, Roslin, Midlothian, Edinburgh, EH25 9PS, UK.

出版信息

BMC Genomics. 2006 Sep 1;7:224. doi: 10.1186/1471-2164-7-224.

DOI:10.1186/1471-2164-7-224
PMID:16948847
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1590031/
Abstract

BACKGROUND

The response of macrophages to danger signals is an important early stage in the immune response. Our understanding of this complex event has been furthered by microarray analysis, which allows the simultaneous investigation of the expression of large numbers of genes. However, the microarray resources available to study these events in livestock animals are limited.

RESULTS

Here we report the development of a bovine macrophage specific (BoMP) cDNA microarray. The BoMP microarray contains 5026 sequence elements (printed in duplicate) and numerous controls. The majority of the clones incorporated on the microarray were derived from the BoMP cDNA library generated from bovine myeloid cells subjected to various stimuli, including over 900 sequences unique to the library. Additional clones representing immunologically important genes have been included on the BoMP microarray. The microarray was validated by investigating the response of bovine monocytes to stimulation with interferon-gamma and lipopolysaccharide using amplified RNA. At 2 and 16 hours post stimulation 695 genes exhibited statistically significant differential expression, including; 26 sequences unique to the BoMP library, interleukin 6, prion protein and toll-like receptor 4.

CONCLUSION

A 5 K cDNA microarray has been successfully developed to investigate gene expression in bovine myeloid cells. The BoMP microarray is available from the ARK-Genomics Centre for Functional Genomics in Farm Animals, UK.

摘要

背景

巨噬细胞对危险信号的反应是免疫反应的重要早期阶段。微阵列分析推动了我们对这一复杂事件的理解，该分析能够同时研究大量基因的表达。然而，可用于研究家畜动物这些事件的微阵列资源有限。

结果

在此我们报告一种牛巨噬细胞特异性（BoMP）cDNA微阵列的开发。BoMP微阵列包含5026个序列元件（一式两份打印）以及众多对照。微阵列上纳入的大多数克隆源自于受各种刺激的牛骨髓细胞所产生的BoMP cDNA文库，包括该文库特有的900多个序列。代表免疫重要基因的其他克隆也已包含在BoMP微阵列上。通过使用扩增RNA研究牛单核细胞对干扰素-γ和脂多糖刺激的反应来验证该微阵列。在刺激后2小时和16小时，695个基因表现出统计学上显著的差异表达，包括：BoMP文库特有的26个序列、白细胞介素6、朊病毒蛋白和Toll样受体4。

结论

已成功开发出一种5K cDNA微阵列来研究牛骨髓细胞中的基因表达。BoMP微阵列可从英国农场动物功能基因组学ARK基因组学中心获得。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f055/1590031/ba0c06ca40b4/1471-2164-7-224-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f055/1590031/d57c9e3eb67f/1471-2164-7-224-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f055/1590031/ba0c06ca40b4/1471-2164-7-224-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f055/1590031/d57c9e3eb67f/1471-2164-7-224-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f055/1590031/ba0c06ca40b4/1471-2164-7-224-2.jpg

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