RESUMO
In this study, we identified myogenic regulatory factors (MRFs) and analyzed the correlation between MRFs and meat quality in rainbow trout. The MyoD1a and MyoD1b genes were cloned from rainbow trout using a homology cloning method. Introns 1 and 2 in the MyoD1a and MyoD1b genes were cloned and submitted to GenBank (accession Nos. FJ623462 and FJ793566). Polymorphisms of MyoD1a and MyoD1b genes were analyzed using single-strand conformation polymorphism and sequencing, respectively. Two single nucleotide polymorphisms were detected in the MyoD1 gene, located at 129GâA in exon 1 and 37 GâA in exon 2. The 37 GâA mutation in exon 2 induced the R185K amino acid change in the polypeptide chain. Seven single nucleotide polymorphisms in the MyoD2 gene were detected, including 218TâC, 224TâC, 242AâC, 246TâA, 248TâC, 305TâC, and 329CâT. The 246TâA mutation in exon 1 induced the R83K change in the polypeptide chain. In the S3 fragment, meat quality traits of genotypes AA and AB significantly differed from those of genotype BB (P < 0.05). In the S5 fragment, meat quality traits of the genotypes AA and AC were significantly different from the genotypes BB and BC (P < 0.05). These results indicate that the MyoD1a and MyoD1b genes have an important influence on meat quality or were linked to the major genes in these strains. These genes can be used to control muscle fiber traits in rainbow trout, and the mutations in the S3 and S5 fragments can be used as molecular markers for selecting rainbow trout with better meat quality traits.
Assuntos
Estudos de Associação Genética , Carne , Proteína MyoD/genética , Oncorhynchus mykiss/genética , Animais , Sequência de Bases , Éxons , Genótipo , Oncorhynchus mykiss/crescimento & desenvolvimento , Fenótipo , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Calpastatin, an important protein used to regulate meat quality traits in animals, is encoded by the CAST gene. The aim of the present study was to clone the cDNA sequence of the CAST gene and detect the expression of CAST in the tissues of Cyprinus carpio. The cDNA of the C. carpio CAST gene, amplified using rapid amplification of cDNA ends PCR, is 2834 bp in length (accession No. JX275386), contains a 2634-bp open reading frame, and encodes a protein with 877 amino acid residues. The amino acid sequence of the C. carpio CAST gene was 88, 80, and 59% identical to the sequences observed in grass carp, zebrafish, and other fish, respectively. The C. carpio CAST was observed to contain four conserved domains with 54 serine phosphorylation loci, 28 threonine phosphorylation loci, 1 tyrosine phosphorylation loci, and 6 specific protein kinase C phosphorylation loci. The CAST gene showed widespread expression in different tissues of C. carpio. Surprisingly, the relative expression of the CAST transcript in the muscle and heart tissues of C. carpio was significantly higher than in other tissues (P < 0.01).
Assuntos
Proteínas de Ligação ao Cálcio/biossíntese , Proteínas de Ligação ao Cálcio/genética , Carpas/genética , Carpas/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , Sequência Conservada , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência , Análise de Sequência de DNA , TranscriptomaRESUMO
The insulin-like growth factor 2 receptor gene (IGF2R) encodes a transmembrane protein receptor and acts to sequester and degrade excess circulating insulin-like growth factor 2, which is critical for normal mammalian growth and development. Thus, IGF2R may serve as a candidate gene underlying growth trait in the common carp. In this study, we isolated the intron one of common carp IGF2R and detected the diversity in 3 continuous generations of FFRC strain common carp. A total of 8 loci were detected within this region, which were named in accordance with their location (i.e., Loc84, Loc106, Loc119, Loc130, Loc145, Loc163, Loc167, and Loc265). Loc106, Loc119, and Loc145 were moderately polymorphic; while Loc84, Loc130, Loc163, Loc167, and Loc265 exhibited slight level of polymorphism. However, significant differences between polymorphism information content values were not observed among the different generations. For Loc145, all generations deviated from Hardy-Weinberg equilibrium. The total number of significant linkage disequilibria for all generations equaled 40. Among them, 4 pairs were detected in each population, while 8 pairs were found in the 2nd and 3rd generations. For Loc130, the G/T genotype exhibited higher body weight when compared to that of the G/G genotype. The frequency of the homozygous G/G genotype reached 87.96%; thus, we can improve FFRC strain common carp growth performance by increasing the percentage of the G/T genotype within a breeding population. Therefore, the G/T genotype could be used as a molecular marker for superior growth traits.
Assuntos
Carpas/crescimento & desenvolvimento , Carpas/genética , Íntrons/genética , Polimorfismo de Nucleotídeo Único/genética , Receptor IGF Tipo 2/genética , Animais , Peso Corporal/genética , Loci Gênicos , Heterozigoto , Desequilíbrio de Ligação/genética , Filogenia , Reação em Cadeia da PolimeraseAssuntos
Linfoma Anaplásico de Células Grandes/patologia , Pâncreas/patologia , Neoplasias Pancreáticas/diagnóstico , Pancreatite/diagnóstico , Neoplasias Retroperitoneais/patologia , Biópsia , Diagnóstico Diferencial , Humanos , Masculino , Invasividade Neoplásica , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/fisiopatologia , Neoplasias Pancreáticas/secundário , Pancreatite/fisiopatologia , Estômago/patologia , Neoplasias Gástricas/patologia , Neoplasias Gástricas/fisiopatologia , Neoplasias Gástricas/secundário , Tomografia Computadorizada por Raios X , Adulto JovemRESUMO
To screen the nucleic acid aptamers of the EB virus-positive nasopharyngeal carcinoma cells, we used SELEX technology and synthesized in vitro a 78-nucleotide random DNA library. We used normal nasopharyngeal epithelial cells and EB virus-positive low differentiated nasopharyngeal carcinoma cells as target to conduct 10 cycles of screening, cloning, sequencing, and identification of the aptamers. The fluorescence produced by the combination of the sub-library and the target cells gained intensity gradually with the increase in the number of screening cycles, indicating elevated binding capacity. The cluster analysis showed that the aptamers can be divided into three families, with two of the families having the common conserved sequence. In this study, by screening nucleic acid aptamers for affinity and specificity, we established an initial aptamer library for EB virus-positive nasopharyngeal carcinoma cells.
Assuntos
Aptâmeros de Nucleotídeos/genética , Neoplasias Nasofaríngeas/genética , Técnica de Seleção de Aptâmeros , Sequência de Bases , Carcinoma , Linhagem Celular Tumoral , Infecções por Vírus Epstein-Barr/genética , Biblioteca Gênica , Herpesvirus Humano 4 , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/virologia , Análise de Sequência de DNARESUMO
Fifty-five Chinese isolates from nodules of Amorpha fruticosa were characterized and compared with the type strains of the species and genera of bacteria which form nitrogen-fixing symbioses with leguminous host plants. A polyphasic approach, which included RFLP of PCR-amplified 16S rRNA genes, multilocus enzyme electrophoresis (MLEE), DNA-DNA hybridization, 16S rRNA gene sequencing, electrophoretic plasmid profiles, cross-nodulation and a phenotypic study, was used in the comparative analysis. The isolates originated from several different sites in China and they varied in their phenotypic and genetic characteristics. The majority of the isolates had moderate to slow growth rates, produced acid on YMA and harboured a 930 kb symbiotic plasmid (pSym). Five different RFLP patterns were identified among the 16S rRNA genes of all the isolates. Isolates grouped by PCR-RFLP of the 16S rRNA genes were also separated into groups by variation in MLEE profiles and by DNA-DNA hybridization. A representative isolate from each of these DNA homology groups had a separate position in a phylogenetic tree as determined from sequencing analysis of the 16S rRNA genes. A new species, Mesorhizobium amorphae, is proposed for the majority of the isolates, which belonged to a moderately slow- to slow-growing, acid-producing group based upon their distinct phylogenetic position, their unique electrophoretic type, their low DNA homology with reference strains representing the species within the genus Mesorhizobium and their distinct phenotypic features. Strain ACCC 19665 was chosen as the type strain for M. amorphae sp. nov.
Assuntos
Rhizobiaceae/classificação , Microbiologia do Solo , Composição de Bases , Sequência de Bases , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Filogenia , Plasmídeos , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 16S/genética , Rhizobiaceae/genéticaRESUMO
The nitrogen-fixing rhizobial symbionts of Sesbania herbacea growing in the nature reserve at the Sierra de Huautla, Mexico, were isolated and characterized. All 104 isolates together with the type strain for Rhizobium galegae, HAMBI 540T, had similar 16S rRNA genes as revealed by PCR-RFLP analysis. Similarity in the sequences of the 16S rRNA genes placed the isolates on a phylogenetic branch shared with R. galegae. Among 66 randomly selected isolates, three closely related electrophoretic alloenzyme types (ETs) were identified, which were distinct from 10 ETs distinguished among 23 strains of R. galegae. A new species Rhizobium huautlense, represented by the Sesbania isolate SO2T, is proposed based upon low estimates of DNA relatedness between our chosen type strain and the type strains for the other species, the dissimilarity of the nucleotide sequence of the 16S rRNA genes, and their distinct ETs compared with R. galegae. The description of R. huautlense is significant because in the reconstruction of the phylogeny at R. huautlense there was a shift in the node of the branch of Agrobacterium vitis relative to that of R. galegae. The revised phylogenetic tree would tend to indicate common ancestry between R. galegae and Rhizobium leguminosarum.