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| Cloning and Expression Analysis of E-Cadherin Gene of Nile Tilapia Oreochromis niloticus |
| YANG Huiyuan1, CHEN Honglin2, LI Ruixue1, YANG Zhihui3, WU Yingrui1, WU Xinzhong1, CAI Xiaohui1 |
1. Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, Beibu Gulf University, Qinzhou 536011, China;
2. College of Life Science and Technology, Guangxi University, Nanning 530004, China;
3. Xingtai Bureau of Agriculture and Rural Affairs, Xingtai 054000, China |
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Abstract The E-cadherin gene of Nile tilapia Oreochromis niloticus (GenBank number: MT370500; named as On-ECdh gene) was cloned, and the bioinformatics characteristics of On-ECdh gene and its expression pattern were analyzed in different tissues of Nile tilapia under normal conditions and stimulation of Streptococcus agalactiae. The specific primers were designed to amplify On-ECdh gene. The characteristics of On-ECdh gene was analyzed by bioinformatics, and its expression pattern was detected by the fluorescence quantitative PCR. The On-ECdh gene had open reading frame of 2442 bp, encoding 813 amino acids. The theoretical molecular weight of On-ECdh protein was 89.9 ku and the isoelectric point was 4.81. Prediction of conservative domain showed that On-ECdh amino acids contained 1 cadherin pro domain, 4 cadherin re-like domain, 1 cadherin cytoplasmic region and 1 transmembrane domain. Homologous alignment showed that On-ECdh amino acids had the highest similarity with Haplochromis burtoni (79.04%), and lower similarity with other fish (42.44%—52.59%) and mammals (37.12%—44.62%). Fluorescence quantitative PCR analysis showed that On-ECdh gene was detected in all the examined tissues of healthy Nile tilapia, with the maximal expression level in liver, followed by gill, intestine, stomach, eye, muscle, spleen, brain, heart, head and kidney. After the stimulation of S. agalactiae, the expression of On-ECdh gene was significantly up-regulated in the head kidneys, eyes and intestines, while down-regulated in the stomach, gills and liver. The expression of On-ECdh gene reached peak at 1 h in eye, 3 h in head kidney and 12 h in intestine, and showed a downward trend after the peak. And its expression levels were the minimum at 1 h in stomach, 3 h in liver and 24 h in gills. Moreover, the expression level of On-ECdh gene showed a time-dependent wavy trend in the stomach and gills, while its expression level in the liver was continuously decreased. The findings showed that S. agalactiae induced the expression of On-ECdh gene, indicating that the On-ECdh gene may play a defensive role in Nile tilapia infected with S. agalactiae.
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Received: 18 May 2020
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