Ten novel polymorphic microsatellite loci were developed using an enrichment and colony hybridization protocol and characterized for the cuttlefish,Sepiella maindroni.Polymorphism was explored by using 30 individuals ...Ten novel polymorphic microsatellite loci were developed using an enrichment and colony hybridization protocol and characterized for the cuttlefish,Sepiella maindroni.Polymorphism was explored by using 30 individuals from the coastal waters of Zhanjiang,Guangdong province,China,in December 2007.The number of alleles ranged from 5 to 13.The ranges of observed and expected heterozygosity were from 0.578 9 to 1.000 0 and 0.682 8 to 0.925 7,respectively,and the average polymorphic information content （P IC） was 0.778 5.These microsatellite loci will certainly facilitate the detection of the genetic variation and population structure of S.maindroni.展开更多
Neuropeptide Y(NPY) has a pivotal role in the regulation of many physiological processes. In this study, the gene encoding a NPY receptor-like from the common Chinese cuttlefish Sepiella japonica( SjNPYR-like) was ide...Neuropeptide Y(NPY) has a pivotal role in the regulation of many physiological processes. In this study, the gene encoding a NPY receptor-like from the common Chinese cuttlefish Sepiella japonica( SjNPYR-like) was identified and characterized. The full-length SjNPYR-like cDNA was cloned containing a 492-bp of 5′ untranslated region(UTR), 1 182 bp open reading frame(ORF) encoding a protein of 393 amino acid residues, and 228 bp of 3′ UTR. The putative protein was predicted to have a molecular weight of 45.54 kDa and an isoelectric point(pI) of 8.13. By informatic analyses, SjNPYR-like was identified as belonging to the class A G protein coupled receptor(GPCR) family(the rhodopsin-type). The amino acid sequence contained 12 potential phosphorylation sites and five predicted N-linked glycosylation sites. Multiple sequence alignment and 3D structure modeling were conducted to clarify SjNPYR bioinformatics characteristics. Phylogenetic analysis identifies it as an NPYR with identity of 33% to Lymnaea stagnalis NPFR. Transmembrane properties of SjNPYR-like were demonstrated in vitro using HEK293 cells and the p EGFP-N1 plasmid. Relative quantifi cation of SjNPYR-like mRNA level confi rmed a high level expression and broad distribution of SjNPYR-like in various tissues of female S. japonica. In addition, the transcriptional profile of SjNPYR-like in the brain, liver, and ovary during gonadal development was analyzed. The results provide basic understanding on the molecular characteristics of SjNPYR-like and its potentially physical functions.展开更多
To explore the effect of environment conditions on immune activity of fish, eight immune-associated genes responsible for innate immunity were selected from the Gen Bank, i.e. Pgrn-a, Ifit2, P-hepcidin, Lect2, β2m, I...To explore the effect of environment conditions on immune activity of fish, eight immune-associated genes responsible for innate immunity were selected from the Gen Bank, i.e. Pgrn-a, Ifit2, P-hepcidin, Lect2, β2m, Irf1, Il25 and Hsp96, and the m RNA expressions of them in the kidney of cultured large yellow croaker Larimichthys crocea in different sea areas in the East China Sea were examined with q PCR techniques. In the contrasts of immune-associated gene expression between areas and populations, significant differences were found, expression levels of these immune-associated genes were lower in the clear water area than in the poor water quantity area, and lower in May than in October. MY was more sensitive to environmental factors than DQ, which was coincident with the water quality in the culturing areas. Differential analyses of the expression levels of these immune-associated genes showed that significant up-regulation could be triggered by poor environmental factors. The expression patterns indicated that the expression levels of these genes were sensitive to ecological changes, thereby the immune-associated genes, especially Pgrn-a, Ifit2, β2m, Il25 and Hsp96, might serve as immediate and sensitive indicators of population immunologic vigor and ecosystem health. But the expression of immunity-associated genes at the level of gene transcription is highly influenced by multiple factors, and the exact causes or influencing factors of the up-regulation or down-regulation of these genes still need further thorough investigation.展开更多
基金The National Key Technology R & D Program of China under contract No.2007BAD43B05the Key Science & Technology Specific Projects of Zhejiang Province under contract No.2007C12062the National Natural Science Foundation of China under contract No.31001109
文摘Ten novel polymorphic microsatellite loci were developed using an enrichment and colony hybridization protocol and characterized for the cuttlefish,Sepiella maindroni.Polymorphism was explored by using 30 individuals from the coastal waters of Zhanjiang,Guangdong province,China,in December 2007.The number of alleles ranged from 5 to 13.The ranges of observed and expected heterozygosity were from 0.578 9 to 1.000 0 and 0.682 8 to 0.925 7,respectively,and the average polymorphic information content （P IC） was 0.778 5.These microsatellite loci will certainly facilitate the detection of the genetic variation and population structure of S.maindroni.
基金Supported by the Public Welfare Technical Applied Research Project of Zhejiang Province(No.2017C32074)the International Science&Technology Cooperation Program of China(No.2014DFT30120)the Open Foundation from Marine Sciences in the Most Important Subjects of Zhejiang(No.20130202)
文摘Neuropeptide Y(NPY) has a pivotal role in the regulation of many physiological processes. In this study, the gene encoding a NPY receptor-like from the common Chinese cuttlefish Sepiella japonica( SjNPYR-like) was identified and characterized. The full-length SjNPYR-like cDNA was cloned containing a 492-bp of 5′ untranslated region(UTR), 1 182 bp open reading frame(ORF) encoding a protein of 393 amino acid residues, and 228 bp of 3′ UTR. The putative protein was predicted to have a molecular weight of 45.54 kDa and an isoelectric point(pI) of 8.13. By informatic analyses, SjNPYR-like was identified as belonging to the class A G protein coupled receptor(GPCR) family(the rhodopsin-type). The amino acid sequence contained 12 potential phosphorylation sites and five predicted N-linked glycosylation sites. Multiple sequence alignment and 3D structure modeling were conducted to clarify SjNPYR bioinformatics characteristics. Phylogenetic analysis identifies it as an NPYR with identity of 33% to Lymnaea stagnalis NPFR. Transmembrane properties of SjNPYR-like were demonstrated in vitro using HEK293 cells and the p EGFP-N1 plasmid. Relative quantifi cation of SjNPYR-like mRNA level confi rmed a high level expression and broad distribution of SjNPYR-like in various tissues of female S. japonica. In addition, the transcriptional profile of SjNPYR-like in the brain, liver, and ovary during gonadal development was analyzed. The results provide basic understanding on the molecular characteristics of SjNPYR-like and its potentially physical functions.
基金provided by The National High Technology Research and Development Program of China (863 Program) (No.2012BAB16B02)
文摘To explore the effect of environment conditions on immune activity of fish, eight immune-associated genes responsible for innate immunity were selected from the Gen Bank, i.e. Pgrn-a, Ifit2, P-hepcidin, Lect2, β2m, Irf1, Il25 and Hsp96, and the m RNA expressions of them in the kidney of cultured large yellow croaker Larimichthys crocea in different sea areas in the East China Sea were examined with q PCR techniques. In the contrasts of immune-associated gene expression between areas and populations, significant differences were found, expression levels of these immune-associated genes were lower in the clear water area than in the poor water quantity area, and lower in May than in October. MY was more sensitive to environmental factors than DQ, which was coincident with the water quality in the culturing areas. Differential analyses of the expression levels of these immune-associated genes showed that significant up-regulation could be triggered by poor environmental factors. The expression patterns indicated that the expression levels of these genes were sensitive to ecological changes, thereby the immune-associated genes, especially Pgrn-a, Ifit2, β2m, Il25 and Hsp96, might serve as immediate and sensitive indicators of population immunologic vigor and ecosystem health. But the expression of immunity-associated genes at the level of gene transcription is highly influenced by multiple factors, and the exact causes or influencing factors of the up-regulation or down-regulation of these genes still need further thorough investigation.
