Cloning and expression analysis of Nrf2 gene in pearl oyster Pinctada fucata martensii exposed to different immune stimulation and nutritional conditions
CAI Caixia, LIAO Yongshan, YANG Chuangye, ZHENG Zhe, DENG Yuewen, WANG Qingheng
1.College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China; 2.Pearl Breeding and Processing Engineering Technology Research Centre of Guangdong Province, Zhanjiang 524088,China; 3.Guangdong Provincial Engineering Laboratory for Mariculture Organism Breeding, Zhanjiang 524088, China
Abstract: To explore the role of nuclear factor erythroid 2 related factor 2(Nrf2)gene in pearl oyster Pinctadafucatamartensii, the complete CDS region of Nrf2 gene was cloned, sequence characteristics and the expression pattern of Nrf2 were detected in different tissues of the pearl oyster by qRT-PCR technology.Then, the expression trends of Nrf2 gene were analyzed in the pearl oysters injected with 100 μL of 10 μg/mL phosphate buffer solution(PBS)(control group), and 100 μL of 10 μg/mL lipopolysaccharide(LPS), peptidoglycan(PNG), and polyinosinic acid(Poly I:C), respectively, and fed isonitrogen and isolipid diets containing different protein sources including Chlorella sp.powder, Spirulinaplatensis powder, yeast powder, soybean meal and corn gluten powder.The Nrf2 gene contained an open reading frame of 2 652 bp, and encoded a total of 883 amino acids with relative molecular mass of 100 150.There was a BRLZ domain in the C-terminal of Nrf2 amino acids.Phylogenetic analysis revealed that Nrf2 of pearl oyster was clustered with other bivalves like Crassostreagigas.In addition, qRT-PCR analysis showed that Nrf2 gene widely existed in tissues including intestine, foot, gonad, mantle, gill, adductor muscle, and hepatopancreas.The expression level of Nrf2 was significantly upregulated under different stimulation including LPS, PGN and Poly I:C at 48 h(P<0.05), which was 3.8, 2.8 and 3.1 times higher than that in the control group at 0 h, respectively.Expression level of Nrf2 was significantly higher in the pearl oyster fed the diets containing Spirulinaplatensis powder, yeast powder and soybean meal diets than that diets containing Chlorella sp.powder and corn gluten powder(P<0.05).The findings indicate that Nrf2 may be involved in the immune response of the pearl oyster, and provide theoretical knowledge for the analysis of the biological functions of Nrf2 and its role in immune regulation.
蔡彩霞, 廖永山, 杨创业, 郑哲, 邓岳文, 王庆恒. 马氏珠母贝Nrf 2基因的克隆及其在不同免疫刺激和营养条件下的表达变化[J]. 大连海洋大学学报, 2022, 37(2): 202-211.
CAI Caixia, LIAO Yongshan, YANG Chuangye, ZHENG Zhe, DENG Yuewen, WANG Qingheng. Cloning and expression analysis of Nrf2 gene in pearl oyster Pinctada fucata martensii exposed to different immune stimulation and nutritional conditions. Journal of Dalian Ocean University, 2022, 37(2): 202-211.
2022, Vol. 37 
