Preparation of ribosomal protein S14 polyclonal antibody in broiler pulmonary artery: Its application in broiler ascites syndrome.
Preparation of ribosomal protein S14 polyclonal antibody in broiler pulmonary artery: Its application in broiler ascites syndrome.
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DOI：
10.1016/j.ijbiomac.2021.10.130
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发表时间：
2021-10
影响因子：
8.2
通讯作者：
Yueming Gu;Xiaoquan Guo;Pei Liu;Guyue Li;Cheng Huang;Fengping Guo;G. Hu;Cong Wu;Zheng Xu;Xianling Yang;Ping Liu
中科院分区：
文献类型：
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作者：
Yueming Gu;Xiaoquan Guo;Pei Liu;Guyue Li;Cheng Huang;Fengping Guo;G. Hu;Cong Wu;Zheng Xu;Xianling Yang;Ping Liu
RPS 14（ribosomal protein S14）基因通过调节核糖体的生物合成和重要蛋白质的翻译来维持机体的正常生理活动。本研究旨在探讨RPS 14在肉鸡腹水综合征（BAS）发病中的作用。我们成功制备了RPS 14的多克隆抗体，并研究了RPS 14蛋白在多种动物关键组织中的定位和表达。本实验利用外源基因的原核表达技术，构建了重组表达质粒PET 28 a-RPS 14。在IPTG诱导条件下，表达了分子量约为22 kDa的His-RPS 14蛋白，纯化后的重组蛋白作为抗原制备兔抗鸡血清。Western blot结果表明，该血清能特异性识别肉鸡重要组织中的RPS 14蛋白。免疫荧光结合同源性分析表明，该抗血清具有明显的种属特异性。与其他物种相比，该蛋白在肉鸡和鸭的关键组织中的表达更为显著。更重要的是，免疫印迹和免疫荧光显示BAS显著降低了RPS 14的表达水平。这进一步表明RPS 14蛋白可以作为BAS研究的重要切入点之一。
RPS14 (ribosomal protein S14) gene maintains the normal physiological activities of the body by regulating the biosynthesis of ribosomes and the translation of important proteins. This study aims to explore the potential role of RPS14 in broiler ascites syndrome (BAS). We successfully prepared polyclonal antibody against RPS14 and studied the localization and expression of RPS14 protein in a variety of animal key tissues. In this experiment, the recombinant expression plasmid PET28a-RPS14 was constructed using the prokaryotic expression technology of foreign genes. Under the conditions of IPTG induction, a His-RPS14 protein with a molecular weight of about 22 kDa was expressed, and the purified recombinant protein was used as an antigen to prepare rabbit anti-chicken serum. Western blot results showed that the serum could specifically identify RPS14 protein in important tissues of broilers. Immunofluorescence combined with homology analysis showed that the antiserum had significant species specificity. Compared with other species, the expression of this protein in key tissues of broilers and ducks was more significant. More importantly, western blotting and immunofluorescence showed that BAS significantly reduced the expression level of RPS14. This further indicated that RPS14 protein can be used as one of the important entry points for BAS research.