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首页> 外文期刊>Journal of Veterinary Research >Expression of vascular cell adhesion molecule 1 (VCAM-1) in the mammary lymph nodes of cows with subclinical mastitis
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Expression of vascular cell adhesion molecule 1 (VCAM-1) in the mammary lymph nodes of cows with subclinical mastitis

机译：患有亚临床乳腺炎的乳腺淋巴结中血管细胞粘附分子1（VCAM-1）的表达

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Introduction: Vascular cell adhesion molecule 1 (VCAM-1) is a member of Ig superfamily. The aim of this study was to prepare highly specific polyclonal antibodies against bovine VCAM-1 and to evaluate the expression of VCAM-1 in the mammary lymph nodes of cows with subclinical mastitis. Material and Methods: The VCAM-1 gene was cloned from bovine Peyer's patches and inserted into the pGEX-4T-1 and pET-28a vectors. The recombinant plasmids pGEX-4T-1/VCAM-1 and pET-28a/VCAM-1 were transferred into Escherichia coli BL21 and the recombinant strains were induced by isopropyl-D-thiogalactoside to produce fusion proteins tagged with polyhistidine (His) and glutathione S-transferase (GST), respectively. The expressed fusion proteins His-VCAM-1 and GST-VCAM-1 were identified by SDS-PAGE and Western blot. His-VCAM-1 protein was used as an antigen to immunise Wistar rats and polyclonal antibody serum against VCAM-1 was obtained. Results: The serum titre tested by indirect ELISA was 128,000 using GST-VCAM-1 as the well coating antigen. Western blots indicated that the antibody recognised recombinant VCAM-1 protein as well as endogenous VCAM-1. In addition, using qPCR and Western blot, VCAM-1 mRNA and protein expression levels were measured in dairy cows with subclinical mastitis. It was demonstrated that VCAM-1 levels in the mammary lymph nodes of the cows were significantly higher than those from healthy controls (P < 0.05). Conclusion: These results are to our knowledge the first report that VCAM-1 expression in the mammary lymph nodes is elevated in dairy cows with subclinical mastitis.
机译：简介：血管细胞粘附分子1（VCAM-1）是IG超家族的成员。该研究的目的是制备对牛VCAM-1的高度特异性多克隆抗体，并评估亚临床乳腺炎的乳腺淋巴结中VCAM-1的表达。材料和方法：VCAM-1基因从牛Peyer的贴片中克隆并插入PGEx-4T-1和PET-28A载体中。将重组质粒PGEX-4T-1 / VCAM-1和PET-28A / VCAM-1转移到大肠杆菌BL21中，并通过异丙基-D-Thiogalactoside诱导重组菌株，以产生标记的融合蛋白，标记为多亚胺（他的）和谷胱甘肽S-转移酶（GST）分别。表达的融合蛋白HIS-VCAM-1和GST-VCAM-1由SDS-PAGE和Western Blot识别。 His-Vcam-1蛋白被用作免疫Wistar大鼠的抗原，并获得对Vcam-1的多克隆抗体血清。结果：间接ELISA测试的血清滴度使用GST-VCAM-1作为井涂层抗原为128,000。 Western印迹表明，抗体识别重组VCAM-1蛋白以及内源Vcam-1。此外，使用QPCR和Western印迹，在具有亚临床乳腺炎的乳制奶牛中测量VCAM-1 mRNA和蛋白表达水平。据证明，奶牛的乳腺淋巴结中的VCAM-1水平明显高于来自健康对照的节点（P <0.05）。结论：这些结果是我们了解乳制睫毛中VCAM-1表达的第一份报告，乳制奶牛患者具有亚临床乳腺炎。

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