Preparation of a JAZF1 protein polyclonal antibody and its potential role in broiler ascites syndrome

Abstract

As a novel functional protein, juxtaposed with another zinc finger protein 1 (JAZF1) can regulate the growth and apoptosis through various pathways, and maintain the body's normal physiological metabolism. To explore the important role of JAZF1 in broiler ascites syndrome (BAS), we analysed the expression and distribution of the protein in poultry and mammal tissues based on the prepared polyclonal antibody. In this study, the recombinant plasmid PET32a-JAZF1 was constructed by TA cloning, subcloning and other technical methods, and the fusion protein His-JAZF1 was successfully expressed. After purification, His-JAZF1 was used as the antigen to prepare high-quality chicken-derived antibodies. Subsequently, the results showed that JAZF1 protein in broiler tissues could be specifically recognized by this antibody. Immunofluorescence showed that JAZF1 protein mainly exists in the cytoplasm of pulmonary artery, liver, kidney, heart and lung tissue cells of various animals. The expression of this protein was more obvious in broiler and duck tissues than in mammalian tissues. In addition, western blotting combined with immunofluorescence showed that BAS caused a significant decrease in JAZF1 protein in tissue cells. This effect further indicated that JAZF1 protein was closely related to the occurrence of BAS and provided a new entry point for the functional study of JAZF1 protein.

Keywords: Broiler ascites syndrome; Juxtaposed with another zinc finger protein 1; Localization analysis; Polyclonal antibody preparation.