Cloning and Expression Analysis of Interferon Regulatory Factor 3（IRF3）Gene in Hybrid Grouper

doi:10.13560/j.cnki.biotech.bull.1985.2019-0324

Abstract

Abstract: Interferon regulatory factor（IRF）family has antiviral and immunoregulatory functions. Interferon regulatory factor 3（IRF-3）is a member of the IRF family and is also an immune-related factor. To understand the immune response of gene IRF3 in the hybrid grouper（Epinephelus fuscoguttatus,♀）×（E. polyphekadion,♂）to exogenous virus stimulation,the IRF3 gene of the hybrid grouper was cloned by RACE（rapid-amplification of cDNA ends）. The full-length cDNA of this gene is 2 529 bp,including 5' non-coding region（5'-UTR）325 bp,3' non-coding region（3'-UTR）916 bp,open reading frame（ORF）1 377 bp,and encoding 458 amino acids. The amino acid sequence comprises of 3 domains,i.e,one N-terminal DNA binding region（DBD）（1-108 aa）,one C-terminal interferon related region（IAD）（255-435 aa）,and a tryptophan rich region（SRD）（440-450 aa）. Phylogenetic analysis showed that the hybrid grouper's IRF3 and the Japanese sea perch's（Lateolabrax maculatus）IRF3 clustered into one,thus the genetic relationship was close. Real-time quantitative PCR was used to detect the expression of gene IRF3 in 9 tissues of liver,stomach,gill,intestine,spleen of hybrid grouper,etc. as well as the temporal expression of peripheral blood lymphocytes（PBL）. The results showed that gene IRF3 expressed in 9 tissues,and the expression in the liver,stomach and intestine was significantly higher than that in other tissues,and the expression in head and kidney was the lowest. The expression of gene IRF3 in PBL increased gradually after stimulation for 1 h at PolyI：C,and reached the maximum at 4 h（about 9.3 times of the control group）,and gradually decreased after 8 h.

Key words: hybrid grouper, interferon regulatory factor 3, gene cloning, peripheral blood lymphocytes, PolyI：C

LU Xiao-ying, HUANG Bao-song, MA Qian, CHEN Gang, WANG Zhong-liang, HUANG Jian-sheng, ERIC AMENYOGBE, XIE Rui-tao, DENG Wen-xin. Cloning and Expression Analysis of Interferon Regulatory Factor 3（IRF3）Gene in Hybrid Grouper[J]. Biotechnology Bulletin, 2019, 35(10): 144-151.

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