Preparation of polyclonal antibody against phosphatidylethanolamine binding protein 1 recombinant protein and its functional verification in pulmonary hypertension syndrome in broilers
Preparation of polyclonal antibody against phosphatidylethanolamine binding protein 1 recombinant protein and its functional verification in pulmonary hypertension syndrome in broilers
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DOI：
10.1016/j.ijbiomac.2022.05.159
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发表时间：
2022-05-27
影响因子：
8.2
通讯作者：
Liu，Ping
中科院分区：
文献类型：
--
作者：
Li，Qingqing;Gu，Yueming;Liu，Ping
肺动脉高压综合征（PHS）是一种困扰快速生长肉鸡的难以克服的疾病。引起肉鸡肺血管重构和腹水。磷脂酰乙醇胺结合蛋白1（PEBP 1）是一种经典的肿瘤转移抑制因子，在肿瘤转移过程中通过多种信号通路调控血管生成。然而，PEBP 1是否能调节PHS肉鸡肺动脉重构尚未见报道。本研究利用基因工程技术构建了[PEBP 1]-pET 32a原核表达载体，在大肠杆菌中大量表达了重组PEBP 1蛋白，并将其免疫家兔制备了PEBP 1多克隆抗体。Western blot和免疫荧光结果表明PEBP 1在多种动物组织中均有表达。但由于多克隆抗体的种属特异性，PEBP 1蛋白在同源性较高的肉鸡和鸭中的表达水平显著高于其他种属动物。更有趣的是，我们发现PEBP 1蛋白的表达在PHS肉鸡中显著降低。这些研究为进一步探讨肺动脉重构的机制奠定了基础。此外，PEBP 1多克隆抗体的制备为进一步研究PEBP 1在PHS中的作用提供了方便。
Pulmonary hypertension syndrome (PHS) is a disease that is difficult to overcome for fast-growing broilers. It causes pulmonary vascular remodeling and ascites in broilers. As a classical inhibitor of cancer metastasis, phosphatidylethanolamine binding protein 1 (PEBP1) regulates angiogenesis in the process of tumor metastasis through multiple signal pathways. However, whether PEBP1 can regulate pulmonary artery remodeling in broilers with PHS has not been reported. This study constructed the prokaryotic expression vector of [PEBP1]-pET32a by genetic engineering technology, the recombinant PEBP1 protein was expressed in large quantities, and the PEBP1 polyclonal antibody was prepared by immunizing rabbits with the recombinant PEBP1 protein. Western blot and immunofluorescence results showed that PEBP1 was expressed in many kinds of animal tissues. However, due to the species specificity of polyclonal antibodies, the expression level of PEBP1 protein in broilers and ducks with high homology was significantly higher than that in other species of animals. More interestingly, we found that the expression of PEBP1 protein decreased significantly in broilers with PHS. These studies laid a foundation for further exploration of the mechanism of pulmonary artery remodeling. In addition, the PEBP1 polyclonal antibody provided convenience for further study of the role of PEBP1 in PHS.