镉胁迫对黄鳝醛酮还原酶基因表达的影响

doi:10.16378/j.cnki.1003-1111.20288

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Abstract The techniques of homology cloning and anchored PCR were used to clone the aldo-keto reductase gene from swamp eel Monopterus albus (Eakr gene). We obtained the genomic DNA sequence from the liver tissue of swamp eel on basis of designed primers according to the intron sequence. The swamp eel was cultured in aerated water of 15 mg/L CdCl₂.The purpose of the study was to preliminarily investigate the role of Eakr gene in swamp eel under Cd²⁺ stress. To determine whether the recombinant Eakr could confer stress tolerance in vitro, the viability of control and Eakr-expressing Escherichia coli under CdCl₂ stress was compared by spot assay and liquid culture. The results showed that a 4468 bp Eakr cDNA sequence was obtained by rapid-amplification of cDNA ends after the sequence were analyzed. The sequence assembly, and the cDNA contained 1026 bp ORF which translated a putative peptide 341 bp amino acids,74 bp 5'UTR and 195 bp 3'UTR. Eakr gene expression level was detected in brain, blood cells, heart, intestine, kidney, liver, muscle, gonad, skin, spleen and stomach of the swamp eel by fluorescence quantitative PCR, with the maximal expression level in liver(P<0.05). Under Cd²⁺ stress, the expression level of Eakr and Nrf2 genes was increased significantly. At each detection time, the expression level of Eakr gene was significantly different from that in the control group(P<0.001). While the expression level of Nrf2 gene was significantly different at 6 h, 9 h, and 12 h after treatment (P<0.001). The recombinant bacteria and control bacteria were treated with CdCl₂. The survival rate of recombinant strains containing Eakr gene was significantly higher than that in negative control, and it was veny significant at Cd²⁺ concentrations of 1.5 mmol/L and 2.0 mmol/L (P<0.001). The finding provides a reference for further understanding of the role of AKR gene in fish in dealing with oxidative stress induced by heavy metals.

Key words： aldo-keto reductase(AKR) Monopterus albus gene clone tissue expression

Received: 06 January 2021

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	KONG Dan
	LI Wei

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KONG Dan,LI Wei. Analysis of Expression of Aldo-Keto Reductase Gene in Swamp Eel Monopterus albus Exposed to Cadmium[J]. Fisheries Science, 2023, 42(1): 119-127.

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https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.20288 OR https://www.shchkx.com/EN/Y2023/V42/I1/119

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