| For aquatic animals, the study on N-acetyltransferase9,Microfibrillar-associated protein 4(MFAP4) and integrins β is still less. In order to adapt to the continuous improvement of people’s living standard, the scale of aquaculture presents the increasing trend, but the subsequent is farmed animal disease outbreaks. Gene cloning of N-acetyltransferase9,Microfibrillar-associated protein 4(MFAP4) and integrins βresearch became instead of antibiotics, treatment and prevention of aquatic animal diseases of choice. Yellow catfish widely distributed in the ocean of eastern China, and is an important food with economic value in nutrition, nourishing and ornamental. But the outbreak of various diseases in recent years has become the main factors which restrict the development of Pelteobagrus fulvidraco breeding high speed. Pelteobagrus fulvidraco N-acetyltransferase9,Microfibrillar-associated protein 4(MFAP4) and integrins β gene and to study the distribution of the mRNA expression has not reported. This study to research the Pelteobagrus fulvidraco, using the method of molecular biology, the length of the get N-acetyltransferase9,Microfibrillar-associated protein 4(MFAP4) and integrins β gene cDNA cloning and amino acid sequence analysis and prediction of protein, molecular biology characteristics of expectation on fish N-acetyltransferase9,Microfibrillar-associated protein 4(MFAP4) and integrinsβgenes increase understanding, also explore the transformation of the physiological regulation of fish protein and provide evidence. With the relative fluorescence quantitative detection of yellow catfish male and female fish in different tissues ofexpression differences.Type peptide analysis, phylogenetic tree constructed using quantitative techniques relative fluorescence detection and tertiary structure prediction of their encoded proteins.N- acetyltransferase 9 in the tertiary structure prediction encoded proteins. For Pelteobagrus fulvidraco lay the foundation for the further study of the growth and disease resistance.Both mannose-binding lectins(MBLs) and ficolins recognize and bind carbohydrates in pathogens and activate complement leading to opsonization, leukocyte activation, and direct pathogen killing. While MBLs have been reported in many fish species, ficolins do not appear to be present in the Pelteobagrus fulvidraco, despite their importance in invertebrate and higher vertebrate innate immunity. A protein with a similar fibrinogenlike domain, microfibrillar-associated protein 4, MFAP4, is present in fish, albeit with no described immune function. We cloned three MFAP4 genes(1MFAP4, 2MFAP4 and 3MFAP4) from the yellow catfish P. fulvidraco. The homology similarity of MFAP4 was 78% with Astyanax mexicanus of Cypriniformes.The yellow catfish MFAP4 transcripts expression analysis revealed difference of patterns of homeostatic expression among the genes in gill, blood, muscle,gonad, liver, brain, spleen, kidney, heart, intestine from male and female. Expression of the three MFAP4 transcripts showed significant changes in expression as soon as 4 h after infection with either Edwardsiella ictaluri or Flavobacterium columnare with modulation of expression continuing up to 24 h or 7 d following pathogen exposure.Several different tissues and gene-specific patterns were captured and transcript expression changes of >11.45-fold were observed over the course of the bacterial challenges. These information will help us elucidate the functions of individual MFAP4 genes in regards to pathogen recognition, binding, and their larger immune context.The N-acetyltransferase 9(NAT9) is an important reproduction related gene. In this study, we cloned the full-length cDNA sequence of yellow catfish Pelteobagrus fulvidraco NAT9 gene. The P. fulvidraco NAT9 cDNA is 1253 bp in length, including584 non-coding region and 669 bp open reading frame. The NAT9 gene encodes a protein of 222 amino acids which shares high homology with the same gene from four other species: Ietalurus Punetaus(91%), Astyanax mexicanus(83%), Danio rerio(81%)and Lepisosteus oculatus(81%). The physicochemical properties,protein domains,protein secondary structure, and 3D structure of the putative NAT9 proteins were analyzed. The NAT9 protein is 25.7899 kD, 4.86 theoretical isoelectric point and C1124H1759N311O359S13 as revealed bycomputer-assisted analysis. Tissue transcription profile indicated that the NAT9 gene is generally but differentially expressed in the detected tissues including gonads, brain, liver, kidney, spleen, intestine, blood, gills,muscle and heart. Identification the sequence and analysis the transcription profile on multiple tissue of of NAT9 genes will provided initial step towards understanding its biological function in yellow catfish, and will also help the production of yellow catfish.Integrinsβ are highly conserved pleiotropic cytokine.In this study, we presented the bioinformatics analysis of TGF- β 1 the full cDNA consists of 2816 bp and encodes 461 aa.TGF- β 1 the full cDNA consists of 1713 bp and encodes 479 aa.Phylogenetic analysis revealed that Integrinsβ were classified strictly with the same type of proteins. Integrinsβ of Stegastes partitus was classified on the same branch of the phylogenetic two with Integrinsβ of other fish. Real-Time PCR result showed that Integrinsβ mRNA was predominantly expressed in the liver and brain,follow by gill and sleepn of the healthy yellow catfish group.We cloned Secondintegrins β genes(integrins β1and integrins β3) from the yellow catfish, P.fulvidraco.The homology similarity of integrins β was 83% ~87% compared with Danio rerio of Cypriniformes.Expression of two Integrinsβ transcripts showed infection with either columnar Flavobacterium(F.columnare), continued until four hours after modulation of expression continuing up to 3d following pathogen exposure.This information to clarify the individual beta gene function in regards to the integration of pathogen recognition,binding and their greater immune environment. |
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