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Gene Cloning and Expression Analysis of Heart-type Fatty Acid-binding Protein in Acipenser dabryanus |
CHEN Ye-Yu, LIU Ya, LAI Jian-Sheng, SONG Ming-Jiang, GONG Quan* |
Fisheries Institute, Sichuan Academy of Agricultural Sciences, Chengdu 611731 China |
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Abstract Heart-type fatty acid-binding protein (FABP-3) is a member of the lipid-binding protein superfamily, and its main functions are uptaking and transportation of fatty acids. In order to understand the structure of FABP-3 and its potential function in nutrition metabolism and immune regulation of Acipenser dabryanus, the full-length cDNA of A. dabryanus FABP-3 was cloned from the unigenes database by full-length transcriptome sequencing of A. dabryanus. The expression profiles of FABP-3 in different tissues, starvation stress and bacterial infection were analyzed. The full length of FABP-3 was 641 bp (GenBank No. MN064725) which contained 89 bp 5' untranslated region (UTR), 150 bp 3' UTR and 402 bp open reading frame (ORF). The ORF of FABP-3 encoded 133-residue peptide. FABP-3 was predicted to have a lipocalin domain from the 6 to 120 amino acids by analysis of the deduced protein sequence using the SMART program. The results also showed the three-dimensional protein structure model predicted by SWISS-MODEL contained 10 antiparallel β-strands and 2 α-helixes in the lipocalin domain, and the 3 conserved amino acid residues (Arg 107, Arg 127, Tyr129) were also present. The tissue distribution and expression level during starvation stress were further analyzed. Results showed that FABP-3 was ubiquitously expressed in all tested tissues, and was mostly expressed in muscle (P≤0.05). During starvation, the expression level of FABP-3 in the brain, liver and stomach increased first and then decreased, while the expression of FABP-3 in the intestine and muscle declined significantly compared to the control group (P≤0.05). Bacterial challenge in vivo revealed significant changes in the gene expression of FABP-3 at immune-related sites. Striking upregulation of FABP-3 was observed in the hindgut at 12 h post-challenge, with increases by more than 219-fold compared to the control, and the expression of FABP-3 mRNA was decreased at first and remarkably elevated in the gill and spleen at 48 h post-challenge. The results indicated that FABP-3 was involved in biological pathways such as nutrition metabolism and immune response in A. dabryanus and would provide basic information for future studies on nutritional and immunomodulatory molecular mechanisms of the A. dabryanus.
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Received: 08 April 2019
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Corresponding Authors:
*admiral671@163.com
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