Cloning And Expression Pattern Analysis Of Rpl10 And TGF-β1 In Spotted Knifejaw(Oplegnathus Punctatus)

With the great success of China’s economic construction,people’s living standards and styles have been greatly improved,the requirements of aquatic products types and quality are also constantly improving,which makes many aquaculture companies vigorously introduce various kinds of aquatic products,and many new types of aquaculture emerge,and spotted knifejaw is one of them.Spotted knifejaw belongs to the genus Porphyridae in the..order Perciformes in morphological classification.It mainly distributes in the northeastern Pacific Ocean.It belongs to the temperate and tropical fishes in coastal areas of China,Japan and Korea.Because of its delicious meat,rich protein content,has a high nutritional and food value,and because of its beautiful body,beautiful body color,has a certain ornamental value,but also because it is difficult to be fished,known as the King of Rock Fishing,has a certain value of fishing entertainment.All these make spotted knifejaw becoming a very popular aquaculture species in the market,and gradually develop into a new aquaculture species.However,the outbreak of large-scale iridovirus disease in the breeding process has seriously restricted the development of the spotted knifejaw breeding industry,causing huge economic losses to the breeders.In this paper,we cloned and analyzed the immune genes related to the anti-iridovirus of spotted knifejaw and preliminarily analyzed the anti-iridovirus immune response mechanism of spotted knifejaw,hoping to provide a reference for further study of the disease resistance mechanism and disease resistance breeding of spotted knifejaw.1 Cloning and expression pattern analysis of rpl10 in spotted knifejawIridoviridae is an important virus family in aquaculture that is featured for its wide infection spectrum in aquatic animals.Recently,spotted knifejaw iridovirus(SKIV)attracts particular attention as it causes high death rate and severe economic loss.Ribosomal protein L10 gene(rpl10)is widely reported to participate the antiviral process in animal and plant.To illustrate their immune roles in spotted knifejaw,we have cloned the rpl10 and studied its expression patterns in this study.The 3463 bp rpl10 genomic sequence consisted of 6 exons and 5 introns,with 1473 bp transcript encoding a 215 amino-acid protein.Tissue expression analysis revealed the wide distribution of rpl10,while higher expression was observed in immune-related tissues(liver and kidney).Time course expression patterns were further examined in three tissues(spleen,liver,kidney)after SKIV infection.Significant up-regulation after SKIV infection could be observed in spleen and liver but not kidney,and the peak levels occurred at 4 days post infection.This study would facilitate the further investigation on functionality of rpl10 in antiviral defense of spotted knifejaw.2 Molecular Cloning and expression pattern analysis of TGF-β1 in spotted knifejawIridovirus has a wide infection spectrum of aquatic animals,which is harmful to aquaculture.In spotted knifejaw,iridovirus is the major factor that causes large-scale mortality.Iridovirus infection results in high mortality rate and thus limits the development of spotted knifejaw industry.As an important immunoregulatory factor,TGF-β1 plays a vital role in viral immune response.In order to study its role in the process of iridovirus infection,we have cloned this gene and studied its spatiotemporal expression patterns.The total length of TGF-β1 gene is 3157 bp,including 6 exons and 5 introns.The open reading frame length is 1167 bp that encodes 388 amino acids.TGF-β1 is widely distributed in tissues,with high expressed in head kidney,intestine,liver,skin,but relatively low expression in spleen and kidney.To further illustrate the expression patterns of TGF-β1 during viral infection,we have analyzed the expression patterns of TGF-β1 in spleen,liver,kidney,and head kidney at different time points after iridovirus infetion.In head kidney,spleen and liver,the expression of TGF-β1 was increased post infection,while the peak level in spleen and liver appeared at 4 day post infection(dpi),and in the head kidney the peak appeared at 10 dpi.In the kidney,viral infection seemed to downregulate TGF-β1 expression.The highest expression was observed at day 0,then decreased at 4 dpi and reach the lowest level at 7 dpi.At 10 dpi the expression was recovered but still lower than level of day 0.These results suggest that TGF-β1 could respond to iridovirus infection and may play an important role in the process of viral immunity.However,the different expression pattern of TGF-β1 in different tissues after viral infection requires further study.