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| Expression of Chicken (Gallus gallus) PPARα Recombinant Protein and Preparation of Its Antibody |
| ZHAO Xiang, GUO Zhi-Jing, LUO Fang, WEI Zhi-Heng, DONG Xiao-Min, XU Lu, YU Jian-Feng*, GU Zhi-Liang* |
| School of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China |
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Abstract Peroxisome proliferator activated receptor α (PPARα) is an important transcriptional regulator involved in lipid metabolism in animals, such as fatty acid oxidation, lipoprotein assembly and transport, lipid catabolism in the liver. So it is particularly important to study the role of PPARα in the lipid metabolism of chicken (Gallus gallus). In order to obtain its effective antibody, the chicken PPARα recombinant protein expressed in Escherichia coli was used to prepare antiserum in Sprague-Dawley (SD) rats (Rattus norvegicus) immunized with the PPARα protein. The titer of the antibodies was detected by Western blot and ELISA. The results showed that PPARα recombinant protein could be soluble expressed in E. coli induced by 0.06 mmol/L isopropyl-β-D-thiogalactopyranoside (IPTG) at 15 ℃, and the purity of the recombinant protein obtained by Ni2+ affinity chromatography was about 89%. The ELISA results showed that the titer of the prepared PPARα antibodies was approximately 1:512 000. The sensitivity of Western blot detection of recombinant protein by antibodies is about 100~200 pg, and endogenous PPARα protein in chicken liver could also be detected. This study provides a reliable antibody for further analysis of the regulatory function of PPARα on the related genes about lipid metabolism in chickens.
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Received: 07 May 2024
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Corresponding Authors:
* csyjf@cslg.edu.cn; zhilianggu88@hotmail.com
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