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| Cloning of Lateolabrax japonicus Haptoglobin Gene and Its Expression Analysis upon Vibrio harveyi Infection |
| QIAO Dan, DING Fei-Fei, LI Chang-Hong*, CHEN Jiong |
| Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo 315832, China |
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Abstract Haptoglobin (Hp), mainly synthesized by the liver, plays an important role in the acute phase response (APR) induced by infection, trauma, and inflammation. In this study, the full-length cDNA sequence of the Hp gene (LjHp) (GenBank No. MK820673) of the Japanese sea bass (Lateolabrax japonicus) was obtained from the transcriptome sequencing data of Japanese sea bass. The cDNA sequence of LjHp consisted of 2 285 nucleotides, which contained a large open reading frame predicted to encode a precursor protein comprising of 309 amino acids with a molecular weight of approximately 33.82 kD. A predicted signal peptide of 22 amino acids was present in the N-terminal of LjHp. Sequence analysis showed that LjHp had the typical characters of fish Hp, and shared the highest amino acid sequence identity (76.8%) with that of European seabream (Dicentrarchus labrax). Phylogenetic tree analysis confirmed that LjHp fell into the fish Hp cluster, and grouped tightly with swamp eel (Monopterus albus) Hp. LjHp mRNA was mainly expressed in the liver of healthy Japanese sea bass, and its expression increased significantly in liver, spleen and head kidney after infection with Vibrio harveyi (P<0.05). Western blot analysis showed that LjHp was modified by N-glycosylation in the serum of the Japanese sea bass. Moreover, the serum content of LjHp up-regulated significantly after infection with V. harveyi (P<0.05). The above results showed that the mRNA and protein expression of LjHp was closely related to V. harveyi infection, revealing that it might be a positive acute phase protein in fish against bacterial infection. The present study provides a theoretical basis for studying the functions of fish Hp and its molecular mechanism in APR induced by pathogens.
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Received: 05 May 2019
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Corresponding Authors:
lichanghong0716@163.com
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