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Now showing items 1 - 16 of 33

  • Cloning and characterization of the gene forl-amino acid oxidase in hybrid tilapia

    Shen, Yubang   Fu, Gui Hong   Liu, Feng   Yue, Gen Hua  

    Tilapia is the common name for a group of cichlid fishes. Identification of DNA markers significantly associated with important traits in candidate genes may speed up genetic improvement. l-Amino acid oxidase (LAO) plays a crucial role in the innate immune defences of animals. Previously, whether LAO variants were associated with economic traits had not been studied in fish. We characterized the cDNA sequence of the LAO gene of hybrid tilapia (Oreochromis spp.). Its ORF was 1536 bp, encoding a flavoenzyme of 511 amino acids. This gene consisted of seven exons and six introns. Its expression was detected in the intestine, blood, kidney, skin, liver. It was highly expressed in the intestine. After a challenge with a bacterial pathogen, Streptococcus agalactiae, its expression was up-regulated significantly in the liver, intestine and spleen (P < 0.05). We identified one SNP in the genomic sequence of the gene and found that this SNP was associated significantly with body length (P < 0.05), but not with resistance to S. agalactiae. The results of this study suggest that the LAO gene plays an important role in innate immune responses to the bacterial pathogen in tilapia. The investigation of relationship between polymorphism of LAO gene and disease resistance and growth in tilapia showed that one SNP was associated significantly with body length. Further experiments on whether SNPs in the LAO gene are associated with growth in tilapia and other populations could be useful in understanding more functions of the LAO gene.
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  • A multiplex microsatellite PCR method for evaluating genetic diversity in grass carp (Ctenopharyngodon idellus)

    Li, Da   Wang, Shentong   Shen, Yubang   Meng, Xinzhan   Xu, Xiaoyan   Wang, Rongquan   Li, Jiale  

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  • A new haplotype variability in complement C6 is marginally associated with resistance to Aeromonas hydrophila in grass carp

    Shen, Yubang   Zhang, Junbin   Xu, Xiaoyan   Fu, Jianjun   Li, Jiale  

    Aeromonas hydrophila, a widespread bacterium in the aquatic environment, causes haemorrhagic septicemia in fish. In the last decade, the disease has caused mass mortality and tremendous economic loss in cultured grass carp in the mainland China. The complement component C6 is a constituent of a biochemical cascade that serves as a major effector of the human innate and adaptor immunity, and eliminates infected cells. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in the C6 gene and to assess their association with A. hydrophila resistance in grass carp. A resource population consisting of 186 susceptible and 191 resistant grass carp was constructed. The gcC6 genomic sequence is composed of 9292 bp, containing 18 exons and 17 introns. The promoter sequence of gcC6 gene contained several consensus sequences for hepatic-specific transcription factors. We sequenced a total of 9744 bp of the C6 gene from a diverse population of grass carp and identified 8 SNPs that were genotyped in the resource population. Statistical analysis revealed a lack of association between any individual SNPs and resistance to A. hydrophila in grass carp. The SNPs 1214G>A, 1380G>C, 2095A>C and 2167T>C were linked together (r(2)>0.8). The haplotype GCCC generated with these four SNPs was associated marginally with resistance to A. hydrophila in grass carp. These findings suggest a lack of strong association of the C6 polymorphisms with the A. hydrophila resistance in grass carp. (C) 2013 Elsevier Ltd. All rights reserved.
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  • Characterization of two novel gadd45a genes in hybrid tilapia and their responses to the infection of Streptococcus agalactiae

    Shen, Yubang   Ma, Keyi   Liu, Feng   Yue, Gen Hua  

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  • Characterization of the duodenase-1 gene and its associations with resistance to Streptococuus agalactiae in hybrid tilapia (Oreochromis spp.)

    Shen, Yubang   Fu, Gui Hong   Liu, Feng   Yue, Gen Hua  

    Highlights • The duodenase-1 gene was characterized for the first time in fish. • The gene encoded a serine protease with 252 amino acids. • The gene was highly expressed in intestine and kidney, and responded to the infection of Streptococcus agalactiae. • SNP haplotype combinations identified in the duodenase-1 gene were significantly associated with the resistance to S. agalactiae. Abstract Tilapia is a group of cultured teleost fishes whose production is threatened by some diseases. Identification of DNA markers associated with disease resistance in candidate genes may facilitate to accelerate the selection of disease resistance. The gene encoding a duodenase, which can trigger immune response, has not been studied in fish. We characterized the cDNA of duodenase-1 gene of hybrid tilapia. Its ORF is 759 bp, encoding a serine protease of 252 amino acids. This gene consisted of five exons and four introns. Its expression was detected in all 10 tissues examined, and it was highly expressed in the intestine and kidney. After a challenge with the bacterial pathogen, Streptococcus agalactiae, its expression was up-regulated significantly in the intestine, liver and spleen. We identified seven SNPs in the gene and found that four of them were significantly associated with the resistance to S. agalactiae (P < 0.05). The CGTCC haplotype, CAGTC/CGGTC and CGTCC/CGTCC diplotype were significantly associated with the resistance to S. agalactiae (P = 0.00, 0.04 and < 0.0001, respectively). In addition, one SNP was associated significantly with growth traits (P < 0.05). These findings suggest that the duodenase-1 gene plays an important role in the resistance to S. agalactiae in tilapia. The SNP markers in the duodenase-1 gene associated with resistance to the bacterial pathogen, may facilitate the selection of tilapia resistant to the bacterial disease.
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  • miR-21 targets jnk and ccr7 to modulate the inflammatory response of grass carp following bacterial infection

    Tao, Lizhu   Xu, Xiaoyan   Fang, Yuan   Wang, Anqi   Zhou, Fenglin   Shen, Yubang   Li, Jiale  

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  • Population structure, demographic history and local adaptation of the grass carp

    Shen, Yubang   Wang, Le   Fu, Jianjun   Xu, Xiaoyan   Yue, Gen Hua   Li, Jiale  

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  • miR-23a-3p and miR-23a-5p target CiGadd45ab to modulate inflammatory response and apoptosis in grass carp

    Fang, Yuan   Xu, Xiao-yan   Shen, Yubang   Li, Jiale  

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  • miR-148 targets CiGadd45ba and CiGadd45bb to modulate the inflammatory response to bacterial infection in grass carp

    Fang, Yuan   Xu, Xiao-yan   Shen, Yubang   Li, Jiale  

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  • Molecular cloning and functional analysis of Growth arrest and DNA damage-inducible 45 as and ab (Gadd45aa and Gadd45 ab) in Ctenopharyngodon idella

    Fang, Yuan   Xu, Xiao-yan   Shen, Yubang   Li, Jiale  

    The Gadd45aa and Gadd45 ab genes are members of the Gadd45 family, which are critically involved in immunological and apoptosis functions. In this study, we isolated and characterized Gadd45aa and Gadd45 ab cDNA from grass carp (Ctenopharyngodon idella) (designated CiGadd45aa and CiGadd45 ab). The CiGadd45aa and CiGadd45 ab fragments spanned 1272 bp/1248 bp, which contained 474 bp/480 bp open reading frames encoding 157/159 amino acid proteins. BLAST analysis revealed that CiGadd45aa and CiGadd45 ab shared high similarity with known Gadd45a sequences. qRT-PCR analysis showed widespread and abundant expression of CiGadd45aa in gill, intestine, kidney, brain, blood, skin and fin, but low in liver, spleen, head kidney, heart, and muscle. CiGadd45 ab was expressed highly in liver, spleen and blood but at low levels in gill, intestine, kidney, head kidney, heart, brain, skin, muscle, and fin. Following challenge of grass carp with Aeromonas hydrophila, CiGadd45aa and CiGadd45 ab expression was upregulated. In immune-relevant tissues and MAPK family genes (p38, JNK and ERK) were upregulated by CiGadd45aa and CiGadd45 ab overexpression and partly downregulated by interfered in the CIK grass carp kidney cell line. In addition, transcription of the cytokine-encoding il-8 gene was upregulated/downregulated by CiGadd45aa and CiGadd45 ab overexpression and interference. These results suggest that CiGadd45aa and CiGadd45 ab play roles in innate immune responses against A. hydrophila in grass carp.
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  • Cloning, functional analysis, and microRNA-induced negative regulation of growth arrest and DNA damage-inducible 45 γ (Gadd45g) in grass carp (Ctenopharyngodon idella)

    Fang, Yuan   Xu, Xiao-Yan   Zhang, Meng   Bai, Yulin   Zhang, Xue-shu   Shen, Yubang   Li, Jiale  

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  • Complement component Bf/C2b gene mediates immune responses against Aeromonas hydrophila in grass carp Ctenopharyngodon idella

    Dang, Yunfei   Shen, Yubang   Xu, Xiaoyan   Wang, Shentong   Meng, Xinzhan   Zhang, Meng   Lv, Liqun   Wang, Rongquan   Li, Jiale  

    The complement system is a significant component of innate immunity. Here, we identified a Bf/C2 homolog (gcBf/C2b) in grass carp. gcBf/C2b shares a high similarity with Bf/C2b counterparts in other teleosts. gcBf/C2b transcription was widely distributed in different tissues, induced by Aeromonas hydrophila in vivo and in vitro, and affected by lipopolysaccharide and fiagellin stimulation in vitro. In cells over-expressing gcBf/C2b, transcript levels of all components except gcCS were significantly enhanced, and gcBf/C2b, gcIL1 beta, gcTNF-alpha, gcIFN, gcCD59, gcC5aR1, and gcITG beta-2 were significantly upregulated after A. hydrophila challenge or stimulation with bacterial pathogen-associated molecular patterns (PAMPs). However, gcBf/C2b in interference cells down-regulated the transcript levels after A. hydrophila challenge, and gcBf/C2b induced NF-kappa B signaling. These findings indicate the vital role of gcBf/C2b in innate immunity in grass carp.
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  • Polymorphic microsatellite loci for population genetics of the hard shelled mussel, Mytilus coruscus

    Shen, Yubang   Bai, Zhiyi   Guo, Shizhao   Li, Jiale  

    Here we report the isolation and characterization of 14 polymorphic loci isolated from a partial genomic DNA library of hard shelled mussel (Mytilus coruscus) enriched for CA repeats. We tested variability of these microsatellites on 60 unrelated individuals collected in China. All loci were polymorphic. The average allele number was 19.9 per locus, ranging from 5 to 36. The observed heterozygosity was from 0.39 to 0.85 with an average of 0.69, whereas the average expected heterozygosity was 0.82. Nine of the 14 microsatellites conformed to Hardy-Weinberg equilibrium, and inherited independently. These developed microsatellites are a potential powerful tool for studies of population genetics and genome mapping for this important food species.
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  • Characterization of 36 insert/deletion mutations by STR genotyping method in grass carp, Ctenopharyngodon idella

    Zhang, Meng   Shen, Yubang   Xu, Xiaoyan   Wang, Rongquan   Leng, Xiangjun   Li, Jiale  

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  • Genome-wide identification of JNK and p38 gene family in Ctenopharyngodon idella and their expression profiles in response to bacterial challenge

    Xu, Xiaoyan   Tao, Lizhu   Wang, Anqi   Li, Liuyang   Fan, Kun   Shen, Yubang   Li, Jiale  

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  • A new haplotype variability in complement C6 is marginally associated with resistance to Aeromonas hydrophila in grass carp

    Shen, Yubang   Zhang, Junbin   Xu, Xiaoyan   Fu, Jianjun   Li, Jiale  

    Aeromonas hydrophila, a widespread bacterium in the aquatic environment, causes haemorrhagic septicemia in fish. In the last decade, the disease has caused mass mortality and tremendous economic loss in cultured grass carp in the mainland China. The complement component C6 is a constituent of a biochemical cascade that serves as a major effector of the human innate and adaptor immunity, and eliminates infected cells. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in the C6 gene and to assess their association with A. hydrophila resistance in grass carp. A resource population consisting of 186 susceptible and 191 resistant grass carp was constructed. The gcC6 genomic sequence is composed of 9292 bp, containing 18 exons and 17 introns. The promoter sequence of gcC6 gene contained several consensus sequences for hepatic-specific transcription factors. We sequenced a total of 9744 bp of the C6 gene from a diverse population of grass carp and identified 8 SNPs that were genotyped in the resource population. Statistical analysis revealed a lack of association between any individual SNPs and resistance to A. hydrophila in grass carp. The SNPs 1214G>A, 1380G>C, 2095A>C and 2167T>C were linked together (r(2)>0.8). The haplotype GCCC generated with these four SNPs was associated marginally with resistance to A. hydrophila in grass carp. These findings suggest a lack of strong association of the C6 polymorphisms with the A. hydrophila resistance in grass carp. (C) 2013 Elsevier Ltd. All rights reserved.
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