Cloning, Identification and Characterization of Interferon System Genes in Crucian Carp (Carassius auratus L.)

doi:10.7523/j.issn.2095-6134.2004.3.022

Abstract

Abstract:

Interferon (IFN) system is the first line of defense against virus invasion in vertebrate. So far relative little was known concerning IFN system genes other than in mammals. In fish, nearly 40 year evidence available for existence of IFN system mainly lies in the observation that fish and fish cells are much earlier known to produce molecules with IFN like activity as measured by a cell protection test, and that IFN inducible gene, Mx homologue, has been identified in a limited number of fish species in the latest years. Previously we reported that UV inactivated GCHV was able to induce high level of IFN like molecule for establishment of antiviral state in cyprinid fish cell lines, such as crucian carp (Carassius auratus L.) blastulae embryonic (CAB) cells. In the current study, in order to better understand innate immune mechanisms of fish cells responsive to virus infection, an ideal cell model system for identifying fish antiviral relevant genes was firstly established, and on the basis of induction and characterization of CAB IFN, a subtractive cDNA library accumulating an enhanced mRNA level of fish antiviral relevant genes was also constructed. Secondly, a subset of fish IFN system genes and some unidentified ESTs that were found no similarity by BLASTX were retrieved by screening of the library, and expression analysis demonstrated that they were IFN stimulated genes (ISGs) as well. Thirdly, according to the latest research of mammals, 19 full length cDNAs of fish IFN system genes were cloned, including crucian carp IFN, IFN signal transduction factor STAT1, IFN regulatory factor IRF7, putative IFN antiviral effectors Mx1, Mx2, PKR, Viperin, IFI56, and some ISGs with function unknown, including IFI58, ISG15 1, ISG15 2, Gig1, Gig2 and so on. Finally, further studies on the characterization of structure, expression, induction and evolution of some important relevant genes reveal that several mechanisms in fish innate immune following virus infection, including signal recognition, signal transduction and regulation, antiviral pathways, were similar to those in mammals.

Key words: crucian carp blastulae embryonic (CAB) cells, interferon system, antiviral and immune relevant gene, gene cloning, antiviral mechanism

CLC Number:

S941.41

ZHANG YiBing, GUI JianFang. Cloning, Identification and Characterization of Interferon System Genes in Crucian Carp (Carassius auratus L.)[J]. , 2004, 21(3): 418-426.

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