草鱼Ⅱ型细胞因子受体基因CRFB1和CRFB5的克隆、鉴定-微生物学专业论文.docxVIP

Abstra Abstract PAGE PAGE IV Abstract Grass carp is one of the most economically important species in Chinas freshwater aquaculture. In breeding process, the high mortality of viral infection has been tormented aquaculture workers. IFN is a cytokine which has multifunction and various biological activities. The recombinant IFN producted by genetic engineering technique has been widely applied to control viral diseases of human and economic animals. Among them, type I IFN plays a vital role in cell antiviral activity. Similar to the mammalian counterparts, fish type I interferon (IFN) performs its potential biological activities by combining with its corresponding receptor on cell membrane. Fish type I IFN receptor, a kind of enzyme-linked receptor, consists of two subunits. Insight into the grass carp I IFN receptor will help us better understand fish IFN signaling pathway and provide theoretical basis for the healthy breeding of fish. Based on zebrfish type I interferon receptor subunits CRFB1 and CRFB5, we cloned and identified two putative grass carp (Ctenopharyngodon idella) type I interferon receptor subunits (termed CiCRFB1 and CiCRFB5) by homology cloning techniques. The full length cDNA sequences of CiCRFB1 and CiCRFB5 are 2945 bp and 1517 bp respectively. Through the sequences analysis，we found that CiCRFB1 and CiCRFB5 exhibits 74 %, 70 % identity to the known DrCRFB1 (EF014952), DrCRFB5 (EF014955) respectively. Phylogenetic analysis results also showed that CiCRFB1 and CiCRFB5 had higher homology to Danio rerio CRFB1 and CRFB5 respectively compared to the counterparts of the other species. Structurally, both CiCRFB1 and CiCRFB5 are composed of a relatively conserved extracellular domain of about 200 amino acids containing two fibronectin type III motifs connected by a linker. Near the extracellular domain, there is a single transmembrane domain behind it. The intracellular domain of CiCRFB1 (279 aa) is far longer than that of CiCRFB5 (129