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1.
Poult Sci ; 96(4): 1015-1027, 2017 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-28339556

RESUMO

p. major: Immature poults have an inefficient thermoregulatory system, and therefore extreme ambient temperatures can impact their internal body temperature. Satellite cells, the only posthatch myonuclei source, are multipotential stem cells and sensitive to temperature. Selection for faster-growing, high-yielding birds has altered satellite-cell properties. The objective of the current study was to determine how temperature affects adipogenic properties of satellite cells isolated from the pectoralis major ( ) muscle of Randombred Control line ( ) and F line turkeys selected only for increased 16-wk body weight from the RBC2 line. Satellite cells were cultured at 2°C incremental temperatures between 33 and 43°C and compared to cells cultured at the control temperature of 38°C to ascertain temperature effects on lipid accumulation and expression of adipogenic genes: CCAAT/enhancer-binding protein-ß ( ), peroxisome proliferator-activated receptor-γ ( ), and stearoyl-CoA desaturase ( ). During proliferation, the amount of quantifiable lipid in both F and RBC2 satellite cells increased at temperatures above 38°C ( P <  0.01) and decreased at temperatures below 38°C ( P < 0.01). Above 38°C, RBC2 satellite cells had more lipid ( P = 0.02) compared to the F line, whereas there were few differences between lines below 38°C. At 72 h of proliferation, expression of C/EBPß , PPARγ , and SCD decreased ( P ≤  0.02) as temperatures increased from 33 to 43°C in both cell lines. During differentiation expression of C/EBPß increased ( P <  0.01) as temperatures increased from 33 to 43°C in both cell lines. In F line satellite cells, PPARγ expression decreased ( P <  0.01) with increasing temperatures during differentiation, whereas there was no linear trend in RBC2 cells. During differentiation expression of SCD increased as temperatures increased ( P <  0.01) in RBC2 cells, and there was no linear trend within the F line. Results from the current study suggest that environmental temperature can affect p. major satellite cellular fate; however, selection for increased body weight had little impact on these cellular responses.


Assuntos
Expressão Gênica , Metabolismo dos Lipídeos , Músculos Peitorais/metabolismo , Células Satélites de Músculo Esquelético/metabolismo , Seleção Genética , Temperatura Ambiente , Perus/metabolismo , Adipogenia , Animais , Peso Corporal , Masculino , Perus/genética
2.
Poult Sci ; 95(4): 934-47, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26769270

RESUMO

Poultry selected for growth have an inefficient thermoregulatory system and are more sensitive to temperature extremes. Satellite cells are precursors to skeletal muscle and mediate all posthatch muscle growth. Their physiological functions are affected by temperature. The objective of the current study was to determine how temperature affects satellite cells isolated from the pectoralis major (p. major) muscle (breast muscle) of turkeys selected for increased 16 wk body weight (F line) in comparison to a randombred control line (RBC2) from which the F line originated. Pectoralis major muscle satellite cells were thermally challenged by culturing between 33°C and 43°C to analyze the effects of cold and heat on proliferation and differentiation as compared to control temperature of 38°C. Expression levels of myogenic regulatory factors: myogenic differentiation factor 1 (MYOD1) and myogenin (MYOG) were quantified by quantitative polymerase chain reaction (qPCR). At all sampling times, proliferation increased at a linear rate across temperature in both the RBC2 and F lines. Differentiation also increased at a linear rate across temperature from 33 to 41°C at all sampling times in both the F and RBC2 lines. Satellite cells isolated from F line turkeys were more sensitive to both hot and cold temperatures as proliferation and differentiation increased to a greater extent across temperature (33 to 43°C) when compared with the RBC2 line. Expression of MYOD1 and MYOG increased as temperatures increased from 33 to 41°C at all sampling times in both the F and RBC2 lines. These results demonstrate that satellite cell function is sensitive to both cold and hot temperatures and p. major muscle satellite cells from F line turkeys are more sensitive to temperature extremes than RBC2 satellite cells.


Assuntos
Temperatura Alta , Músculos Peitorais/fisiologia , Células Satélites de Músculo Esquelético/fisiologia , Perus/fisiologia , Aclimatação , Animais , Diferenciação Celular , Proliferação de Células , Masculino , Músculos Peitorais/crescimento & desenvolvimento , Seleção Genética , Perus/genética , Perus/crescimento & desenvolvimento
3.
J Anim Sci ; 92(3): 1250-60, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24496837

RESUMO

Previous studies from our laboratory suggested that differential expression of genes between normal and pale, soft, and exudative (PSE) turkey is associated with development of the PSE syndrome. However, a detailed understanding of molecular mechanisms responsible for the development of this meat defect remains unclear. The objective of this study was to extend and complement our previous work by using deep transcriptome RNA sequence analysis to compare the respective transcriptome profiles and identify molecular mechanisms responsible for the etiology of PSE turkey meat. Turkey breasts (n = 43) were previously classified as normal or PSE using marinade uptake as an indicator of quality (high = normal; low = PSE). Total RNA from breast muscle samples with the highest (n = 4) and lowest (n = 4) marinade uptake were isolated and sequenced using the Illumina GA(IIX) platform. The results indicated differential expression of 494 loci (false discovery rate < 0.05). Changes in gene expression were confirmed using quantitative real-time PCR. Pathway analysis of differentially expressed genes suggested abnormalities of calcium homeostasis and signaling pathways regulating actin cytoskeleton structure as well as carbohydrate metabolism and energy production in PSE samples. Dysregulation of postmortem glucose oxidation in PSE turkey was suggested by both dramatic downregulation of pyruvate dehydrogenase kinase, isozyme 4 (PDK4) mRNA, the most downregulated gene, and a decrease in the protein product (P = 0.0007) as determined by immunoblot analysis. These results support the hypothesis that differential expression of several genes and their protein products contribute to development of PSE turkey.


Assuntos
Carne/normas , Transcriptoma/genética , Transcriptoma/fisiologia , Animais , Regulação da Expressão Gênica , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Perus/genética , Perus/metabolismo
4.
Poult Sci ; 92(6): 1523-34, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23687148

RESUMO

The major histocompatibility complex (MHC) is a highly polymorphic region of the genome essential to immune responses and animal health. In galliforms, the MHC is divided into 2 genetically unlinked regions (MHC-B and MHC-Y). Many MHC-B genes are involved in adaptive or innate immunity, yet others have nonimmune or unknown functions. The sequenced MHC-B region of the turkey (Meleagris gallopavo) contains 40 genes, the majority of which are predicted transcripts based on comparison with the chicken or quail, without direct evidence for expression. This study was designed to test for the presence of MHC-B gene transcripts in a panel of immune and nonimmune system tissues from domestic turkeys. This analysis provides the first locus-wide examination of MHC-B gene expression in any avian species. Most MHC-B genes were broadly expressed across tissues. Expression of all predicted genes was verified by reverse-transcription PCR, including B-butyrophilin 2 (BTN2), a predicted gene with no previous evidence for expression in any species. Previously undescribed splice variants were also detected and sequenced from 3 genes. Characterization of MHC-B expression patterns helps elucidate unknown gene functions and potential gene coregulation. Determining turkey MHC-B expression profiles increases our overall understanding of the avian MHC and provides a necessary resource for future research on the immunological response of these genes.


Assuntos
Regulação da Expressão Gênica/fisiologia , Complexo Principal de Histocompatibilidade/genética , Complexo Principal de Histocompatibilidade/fisiologia , Transcriptoma , Perus/genética , Animais , Regulação da Expressão Gênica/imunologia , Regulação da Expressão Gênica no Desenvolvimento , Músculo Esquelético/metabolismo , Reprodutibilidade dos Testes
5.
Poult Sci ; 92(6): 1621-33, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23687159

RESUMO

In response to high consumer demand, turkeys have been intensively selected for rapid growth rate and breast muscle mass and conformation. The success in breeding selection has coincided with an increasing incidence of pale, soft, and exudative (PSE) meat defect, especially in response to heat stress. We hypothesized that the underlying mechanism responsible for the development of PSE meat arises from differences in expression of several critical genes. The objective of this study was to determine differential gene expression between normal and PSE turkey meat using a 6K turkey skeletal muscle long oligonucleotide microarray. Breast meat samples were collected from Randombred Control Line 2 turkeys at 22 wk of age, and classified as normal or PSE primarily based on marinade uptake (high = normal, low = PSE). Total RNA was isolated from meat samples with the highest (normal, n = 6) and the lowest (PSE, n = 6) marinade uptake. Microarray data confirmation was conducted using quantitative real-time PCR. Selection of differentially expressed genes for pathway analysis was performed using a combination of fold change (FC) ranking (FC < -1.66, FC >1.66) and false discovery rate (<0.35) as criteria. The calcium signaling pathway was highlighted as the top canonical pathway associated with differential gene expression between normal and PSE turkey. Dramatic downregulation of fast-twitch myosin heavy chain coupled with upregulation of slow-twitch myosin and troponin C suggested a switch of skeletal muscle isoforms, which may alter muscle fiber arrangement and formation of actin-myosin complexes. Changes in expression of genes in the actin cytoskeleton signaling pathway also suggest altered structures of actin filaments that may affect cell motility as well as strength and flexibility of muscle cells. Substantial downregulation of pyruvate dehydrogenase kinase, isozyme 4 was observed in PSE samples, suggesting altered regulation of the aerobic metabolic pathway in the birds that developed PSE meat defect.


Assuntos
Regulação da Expressão Gênica/fisiologia , Carne/normas , Músculo Esquelético/metabolismo , Actinas/metabolismo , Animais , Transdução de Sinais , Transcriptoma , Perus/genética , Perus/metabolismo , Proteína rhoA de Ligação ao GTP/genética , Proteína rhoA de Ligação ao GTP/metabolismo
6.
Immunogenetics ; 65(2): 133-44, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23179555

RESUMO

The major histocompatibility complex (MHC) plays a central role in innate and adaptive immunity, but relatively little is known about the evolution of the number and arrangement of MHC genes in birds. Insights into the evolution of the MHC in birds can be gained by comparing the genetic architecture of the MHC between closely related species. We used a fosmid DNA library to sequence a 60.9-kb region of the MHC of the greater prairie chicken (Tympanuchus cupido), one of five species of Galliformes with a physically mapped MHC. Greater prairie chickens have the smallest core MHC yet observed in any bird species, and major changes are observed in the number and arrangement of MHC loci. In particular, the greater prairie chicken differs from other Galliformes in the deletion of an important class I antigen binding gene. Analysis of the remaining class IA gene in a population of greater prairie chickens in Wisconsin, USA revealed little evidence for selection at the region responsible for antigen binding.


Assuntos
Genes MHC Classe I , Loci Gênicos , Transportadores de Cassetes de Ligação de ATP/genética , Sequência de Aminoácidos , Animais , Galinhas , Biologia Computacional/métodos , Feminino , Ordem dos Genes , Rearranjo Gênico , Genômica , Dados de Sequência Molecular , Polimorfismo Genético , Alinhamento de Sequência , Homologia de Sequência
7.
Poult Sci ; 91(8): 1964-73, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22802192

RESUMO

Our previous transcriptional profiling study using a turkey skeletal muscle-specific oligonucleotide microarray revealed over 3,000 genes that were differentially expressed at 3 critical stages of muscle development: 18 d embryonic, 1 d posthatch, and 16 wk of age. The genes versican, matrix Gla protein (MGP), and death-associated protein (DAP) were selected to study for their potential effects on muscle satellite cell proliferation and differentiation, as their functions in other tissues are suggestive of possible key roles in the regulation of myogenesis and they are differentially expressed throughout muscle development in the turkey. Using small interfering RNA to knockdown the expression of these genes during proliferation and differentiation, each of the genes was found to differentially affect proliferation and differentiation. Versican and MGP predominantly affected proliferation with line effects, but later stages of differentiation were affected by the knockdown of versican and MGP. The underexpression of DAP inhibited myotube formation, which is a necessary stage in the development of muscle fibers. Without myotube development, muscle fiber formation will be inhibited or abolished. This is the first report that these genes with no previously documented functions with regard to muscle development play a critical role in muscle cell proliferation and differentiation.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica/fisiologia , Proteínas Mitocondriais/metabolismo , Células Satélites de Músculo Esquelético/citologia , Perus/metabolismo , Versicanas/metabolismo , Animais , Proteínas de Ligação ao Cálcio/genética , Diferenciação Celular , Proliferação de Células , Proteínas da Matriz Extracelular/genética , Masculino , Músculo Esquelético/metabolismo , Células Satélites de Músculo Esquelético/metabolismo , Fatores de Tempo , Versicanas/genética
8.
Langmuir ; 28(18): 7291-8, 2012 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-22502638

RESUMO

A series of five near-monodisperse sterically stabilized polystyrene (PS) latexes were synthesized using three well-defined poly(glycerol monomethacrylate) (PGMA) macromonomers with mean degrees of polymerization (DP) of 30, 50, or 70. The surface coverage and grafting density of the PGMA chains on the particle surface were determined using XPS and (1)H NMR spectroscopy, respectively. The wettability of individual latex particles adsorbed at the air-water and n-dodecane-water interfaces was studied using both the gel trapping technique and the film calliper method. The particle equilibrium contact angle at both interfaces is relatively insensitive to the mean DP of the PGMA stabilizer chains. For a fixed stabilizer DP of 30, particle contact angles were only weakly dependent on the particle size. The results are consistent with a model of compact hydrated layers of PGMA stabilizer chains at the particle surface over a wide range of grafting densities. Our approach could be utilized for studying the adsorption behavior of a broader range of sterically stabilized inorganic and polymeric particles of practical importance.


Assuntos
Látex/química , Poliestirenos/química , Molhabilidade , Adsorção , Látex/síntese química , Tamanho da Partícula , Poliestirenos/síntese química , Propriedades de Superfície
9.
Anim Genet ; 43(3): 298-308, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22486501

RESUMO

Skeletal muscle is composed of metabolically heterogeneous myofibres that exhibit high plasticity at both the morphological and transcriptional levels. The objective of this study was to employ microarray analysis to elucidate the differential gene expression between the tonic-'red' anterior latissimus dorsi (ALD) muscle, the phasic-'white' posterior latissimus dorsi (PLD) and 'mixed'-phenotype biceps femoris (BF) in 1-week-and 19-week-old male turkeys. A total of 170 differentially expressed genes were identified in the muscle samples analysed (P < 0.05). Gene GO analysis software was utilized to identify top gene networks and metabolic pathways involving differentially expressed genes. Quantitative real-time PCR for selected genes (BAT2D, CLU, EGFR and LEPROT) was utilized to validate the microarray data. The largest differences were observed between ALD and PLD muscles, in which 32 genes were over-expressed and 82 genes were under-expressed in ALD1-PLD1 comparison, and 70 genes were over-expressed and 70 under-expressed in ALD19-PLD19 comparison. The largest number of genes over-expressed in ALD muscles, as compared to other muscles, code for extracellular matrix proteins such as dystroglycan and collagen. The gene analysis revealed that phenotypically 'red' BF muscle has high expression of glycolytic genes usually associated with the 'white' muscle phenotype. Muscle-specific differences were observed in expression levels of genes coding for proteins involved in mRNA processing and translation regulation, proteosomal degradation, apoptosis and insulin resistance. The current findings may have large implications in muscle-type-related disorders and improvement of muscle quality in agricultural species.


Assuntos
Fibras Musculares Esqueléticas/metabolismo , Proteínas Musculares/metabolismo , Perus/metabolismo , Fatores Etários , Animais , DNA Complementar/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Masculino , Carne , Proteínas Musculares/genética , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Reação em Cadeia da Polimerase em Tempo Real , Perus/genética , Perus/crescimento & desenvolvimento
10.
Cytogenet Genome Res ; 132(1-2): 55-63, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-20733277

RESUMO

Two genetically unlinked gene clusters currently define the turkey major histocompatibility complex (MHC). Previous studies identified turkey bacterial artificial chromosome (BAC) clones hypothesized as orthologs of the MHC-B and MHC-Y regions of the chicken. Physical mapping assigned these clones to the same microchromosome (MGA18) and sequencing of the MHC-B BAC found near synteny with a portion of the chicken B-locus. This study examines the sequence of the second MHC BAC clone that was hypothesized, based on subclone sequences, to be orthologous to the MHC-Y. Sequencing of this clone identified a class I locus and orthologs of additional genes found in the mammalian class III region. Approximately 50% of the BAC insert is comprised of sequence corresponding to the centromeric repeat, MGASat2. This turkey MHC BAC sequence is unique from sequences assigned to the MHC-Y in the chicken. Based on sequence comparisons, the class I gene appears to be a nonfunctional pseudogene. The class III genes (BAT1, BAT3, STK19, and a G4-like locus) represent the second class III gene cluster identified in the galliform genome. This cluster appears to be of ancient origin and provides insight into the evolution of the avian MHC.


Assuntos
Cromossomos Artificiais Bacterianos , Complexo Principal de Histocompatibilidade/genética , Família Multigênica , Perus/genética , Animais , Sequência de Bases , Southern Blotting , DNA , Dados de Sequência Molecular , Homologia de Sequência do Ácido Nucleico
11.
Anim Genet ; 42(1): 75-82, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20528844

RESUMO

Consumer demand for lean, inexpensive meat products has driven the domestic turkey (Meleagris gallopavo) industry to unprecedented production; however, this has coincided with an increase in growth-induced myopathies and meat quality defects. With the aim of developing a new tool for the study of turkey growth and development at the muscle transcriptome level, a 6K oligonucleotide microarray was constructed, the Turkey Skeletal Muscle Long Oligo (TSKMLO) microarray. Skeletal muscle samples were collected at three critical stages in muscle development: 18-day embryo (hyperplasia), 1-day post-hatch (hypertrophy), and 16-week (market age) from two genetic lines of turkeys: RBC2, a line maintained without selection pressure, and F, a line selected from the RBC2 line for increased 16-week body weight. Oligonucleotides were designed from sequences obtained from skeletal muscle cDNA libraries from the three developmental stages. Several unique controls, including mismatch and distance controls and scrambled sequences, were designed for 30 genes. Quality control hybridizations were completed, confirming the validity and repeatability of the array. Control features were evaluated across two larger experiments comparing developmental stage within genetic line or genetic line within each developmental stage, totaling 70 arrays. Mismatch and scrambled sequences appeared to be useful controls of specific hybridization for most genes. In addition, quantitative real-time RT-PCR confirmed microarray results. This creation and assessment of the TSKMLO array provides a valuable community resource for the study of gene expression changes related to turkey muscle growth and development.


Assuntos
Perfilação da Expressão Gênica/veterinária , Carne , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Perus/crescimento & desenvolvimento , Perus/genética , Animais , Biblioteca Gênica , Desenvolvimento Muscular , Músculo Esquelético/crescimento & desenvolvimento , Músculo Esquelético/metabolismo , Doenças Musculares/veterinária , Análise de Sequência com Séries de Oligonucleotídeos/métodos
12.
Heredity (Edinb) ; 107(1): 40-9, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21179065

RESUMO

Studies of major histocompatibility complex (MHC) diversity in non-model vertebrates typically focus on structure and sequence variation in the antigen-presenting loci: the highly variable and polymorphic class I and class IIB genes. Although these studies provide estimates of the number of genes and alleles/locus, they often overlook variation in functionally related and co-inherited genes important in the immune response. This study utilizes the sequence of the MHC B-locus derived from a commercial turkey to investigate MHC variation in wild birds. Sequences were obtained for nine interspersed MHC amplicons (non-class I/II) from each of 40 birds representing 3 subspecies of wild turkey (Meleagris gallopavo). Analysis of aligned sequences identified 238 single-nucleotide variants approximately one-third of which had minor allele frequencies >0.2 in the sampled birds. PHASE analysis identified 70 prospective MHC haplotypes in the wild turkeys, whereas a combined analysis with commercial birds identified almost 100 haplotypes in the species. Denaturing gradient gel electrophoresis (DGGE) of the class IIB loci was used to test the efficacy of single-nucleotide polymorphism (SNP) haplotyping to capture locus-wide variation. Diversity in SNP haplotypes and haplotype sharing among individuals was directly reflected in the DGGE patterns. Utilization of a reference haplotype to sequence interspersed regions of the MHC has significant advantages over other methods of surveying diversity while identifying high-frequency SNPs for genotyping. SNP haplotyping provides a means to identify both divergent haplotypes and homozygous individuals for assessment of immunological variation in wild and domestic populations.


Assuntos
Variação Genética , Complexo Principal de Histocompatibilidade/genética , Perus/genética , Alelos , Animais , Eletroforese em Gel de Gradiente Desnaturante , Loci Gênicos , Genótipo , Haplótipos , Polimorfismo de Nucleotídeo Único
13.
Anim Genet ; 40(6): 925-32, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19744144

RESUMO

The previous genetic mapping data have suggested that most of the rainbow trout sex chromosome pair is pseudoautosomal, with very small X-specific and Y-specific regions. We have prepared an updated genetic and cytogenetic map of the male rainbow trout sex linkage group. Selected sex-linked markers spanning the X chromosome of the female genetic map have been mapped cytogenetically in normal males and genetically in crosses between the OSU female clonal line and four different male clonal lines as well as in outcrosses involving outbred OSU and hybrids between the OSU line and the male clonal lines. The cytogenetic maps of the X and Y chromosomes were very similar to the female genetic map for the X chromosome. Five markers on the male maps are genetically very close to the sex determination locus (SEX), but more widely spaced on the female genetic map and on the cytogenetic map, indicating a large region of suppressed recombination on the Y chromosome surrounding the SEX locus. The male map is greatly extended at the telomere. A BAC clone containing the SCAR (sequence characterized amplified region) Omy-163 marker, which maps close to SEX, was subjected to shotgun sequencing. Two carbonyl reductase genes and a gene homologous to the vertebrate skeletal ryanodine receptor were identified. Carbonyl reductase is a key enzyme involved in production of trout ovarian maturation hormone. This brings the number of type I genes mapped to the sex chromosome to six and has allowed us to identify a region on zebrafish chromosome 10 and medaka chromosome 13 which may be homologous to the distal portion of the long arm of the rainbow trout Y chromosome.


Assuntos
Oncorhynchus mykiss/genética , Recombinação Genética , Cromossomo Y , Animais , Feminino , Masculino
14.
Cytogenet Genome Res ; 125(1): 67-73, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19617698

RESUMO

Cytochromes P450 (P450 for protein; CYP for gene) are a superfamily of membrane-bound hemoproteins that oxidize a large number of endogenous and exogenous compounds. Through oxidation reactions, these enzymes are often responsible for the toxic and carcinogenic effects of natural food-borne toxicants, such as the mycotoxin aflatoxin B1 (AFB1). Previous studies in our laboratory have shown that the extreme sensitivity of turkeys to AFB1 is in part explained by efficient hepatic P450-mediated epoxidation to the toxic and reactive metabolite the exo-AFB1-8,9-epoxide (AFBO). Using 3'-5'-rapid amplification of cDNA ends (RACE), we amplified CYP3A37 from turkey liver RNA, the E. coli-expressed protein which efficiently epoxidates AFB(1). Turkey CYP3A37 has an ORF of 1512 bp, and the protein is predicted to be 504 amino acids with 97% homology to chicken CYP3A37. The turkey gene is organized into 13 exons and 12 introns. A single nucleotide polymorphism in the 11th intron was used to assign CYP3A37 to turkey linkage group 10 (corresponding to chicken chromosome 14, GGA14). Because of the important role of P450s in the extreme sensitivity of turkeys to the toxic effects of AFB(1), this study will contribute to the identifying allelic variants of this important gene in poultry.


Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Perus/genética , Aflatoxina B1/farmacocinética , Aflatoxina B1/toxicidade , Sequência de Aminoácidos , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Sequência de Bases , Galinhas/genética , Mapeamento Cromossômico , Família 3 do Citocromo P450 , Primers do DNA/genética , DNA Complementar/genética , Fígado/metabolismo , Dados de Sequência Molecular , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Perus/metabolismo
15.
Anim Genet ; 40(3): 348-52, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19292710

RESUMO

Vertebrate whole genome sequence assembly can benefit from a priori knowledge of variability in the target genome, with researchers often selecting highly inbred individuals for sequencing. However, for most species highly inbred research lines are lacking, requiring the use of an outbred individual(s). Here we examined the source DNA [Nicholas inbred (Nici)] of the CHORI-260 turkey bacterial artificial chromosome (BAC) library through analysis of microsatellites and BAC sequences. Heterozygosity of Nici was compared with that of individuals from several breeder lines. Seventy-eight microsatellites were screened for polymorphism in a total of 43 birds, identifying an average individual heterozygosity of 0.39, with Nici at 0.35. Additional loci (total of 147) were examined on a subset of individuals to obtain better genome coverage. The mean heterozygosity for this subset was 0.33 with Nici at 0.31. Examination of approximately 200 kb of genome sequence identified SNPs in the order of one per 200 bp in Nici. These data suggest that the heterozygosity of Nici is comparable to other birds of selected breeder lines and that whole genome sequencing would result in an abundant resource of genome-wide polymorphisms.


Assuntos
Cromossomos Artificiais Bacterianos/genética , DNA/genética , Perus/genética , Animais , Mapeamento Cromossômico , DNA/química , Feminino , Variação Genética , Genótipo , Masculino , Repetições de Microssatélites , Polimorfismo de Nucleotídeo Único
16.
Anim Genet ; 39(6): 635-44, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18828862

RESUMO

This study was designed to identify important muscle gene homologues in the turkey. Three skeletal muscle cDNA libraries representing distinct muscle developmental stages were constructed. A total of 20,042 clones were sequenced resulting in 13,023 finished high-quality sequences (trimmed, quality scored and masked) for analysis. Sequence clustering produced 1113 contigs and 4144 singletons (5257 putative transcripts). Sequences were compared by blastn to the chicken whole-genome sequence and to the Ensembl and NCBI databases to identify homologous sequences. These surveys indicated that most of the important muscle genes are included in the sequence collection. Examination of contigs identified 1288 single nucleotide polymorphisms and in 320 of those the minor allele was observed to be present in more than one sequence. This resource provides sequence variants for numerous genes in the turkey, as demonstrated by the SNP haplotypes that were constructed for 10 genes. Sequences obtained in this study provide the basis for constructing a skeletal muscle-focused microarray, a tool that will facilitate the analysis of genes expressed during turkey muscle development, as well as the expression of genes underlying the genetic basis of muscle characteristics associated with meat quality.


Assuntos
Etiquetas de Sequências Expressas , Músculo Esquelético/metabolismo , Perus/genética , Animais , DNA Complementar , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único
17.
Anim Biotechnol ; 19(4): 243-309, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18855250

RESUMO

Trace sequences from the 2X alpaca genome sequencing effort were examined to identify simple sequence repeats (microsatellites) for genetic studies. A total of 6,685 repeat-containing sequences were downloaded from GenBank, processed, and assembled into contigs representing an estimated 4,278 distinct sequences. This sequence set contained 2,290 sequences of length > 100 nucleotides that contained microsatellites of length > or = 14 dinucleotide or 10 trinucleotide repeats with purity equal to 100%. An additional 13 sequences contained a GC microsatellite of length > or = 12 repeats (purity = 100%) were also obtained. Primer pairs for amplification of 1,516 putative loci are presented. Amplification of genomic DNA from alpaca and llama by PCR was demonstrated for 14 primer sets including one from each of the microsatellite repeat types. Comparative chromosomal location for the alpaca markers was predicted in the bovine genome by BLAT searches against assembly 4.0 of the bovine whole genome sequence. A total of 634 markers (41.8%) returned BLAT hits with score > 100 and Identity > 85%, with the majority assignable to unique locations. We show that microsatellites are abundant and easily identified within the alpaca genome sequence. These markers will provide a valuable resource for further genetic studies of the alpaca and related species.


Assuntos
Camelídeos Americanos/genética , Repetições de Microssatélites , Animais , Análise por Conglomerados , DNA/química , DNA/genética , Marcadores Genéticos/genética , Reação em Cadeia da Polimerase , Polimorfismo Genético
18.
Poult Sci ; 87(6): 1165-70, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18493007

RESUMO

Although the domestic turkey (Meleagris gallopavo) is a valuable agricultural commodity, genetic studies on this species lag behind those of other agricultural species. In this study, we examined expressed sequence tags (EST) from a turkey cardiac cDNA library constructed from 4 birds representing 2 developmental stages. A collection of 3,937 EST sequences were sequenced and analyzed for gene annotation and sequence variation. Clustering of sequences resulted in 353 contigs and 874 singletons (1,227 putative transcripts). All EST sequences were compared by BLASTN to the chicken whole genome sequence and to Ensembl and National Center for Biotechnology Information databases. The majority of significant matches correspond to genes found in the chicken. Sequence polymorphisms were identified in 310 contigs, 64 where the minor allele was observed to be present in more than 1 sequence. This study gives species-specific insight into the cardiac transcriptome of turkeys and provides resources for future studies of cardiac function.


Assuntos
DNA Complementar/genética , Biblioteca Gênica , Coração , Perus/genética , Animais , Cardiomiopatia Dilatada/genética , Cardiomiopatia Dilatada/veterinária , Galinhas/genética , Etiquetas de Sequências Expressas , Doenças das Aves Domésticas/genética , Proteínas/genética , RNA/genética , RNA/isolamento & purificação
19.
Cytogenet Genome Res ; 119(1-2): 113-26, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-18160790

RESUMO

An integrated genetic linkage map was developed for the turkey (Meleagris gallopavo) that combines the genetic markers from the three previous mapping efforts. The UMN integrated map includes 613 loci arranged into 41 linkage groups. An additional 105 markers are tentatively placed within linkage groups based on two-point LOD scores and 19 markers remain unlinked. A total of 210 previously unmapped markers has been added to the UMN turkey genetic map. Markers from each of the 20 linkage groups identified in the Roslin map and the 22 linkage groups of the Nte map are incorporated into the new integrated map. Overall map distance contained within the 41 linkage groups is 3,365 cM (sex-averaged) with the largest linkage group (94 loci) measuring 533.1 cM. Average marker interval for the map was 7.86 cM. Sequences of markers included in the new map were compared to the chicken genome sequence by 'BLASTN'. Significant similarity scores were obtained for 95.6% of the turkey sequences encompassing an estimated 91% of the chicken genome. A physical map of the chicken genome based on positions of the turkey sequences was built and 36 of the 41 turkey linkage groups were aligned with the physical map, five linkage groups remain unassigned. Given the close similarities between the turkey and chicken genomes, the chicken genome sequence could serve as a scaffold for a genome sequencing effort in the turkey.


Assuntos
Genoma/genética , Perus/genética , Animais , Sequência de Bases , Galinhas/genética , Mapeamento Cromossômico , Biologia Computacional , DNA/genética , Ligação Genética/genética , Marcadores Genéticos/genética , Família Multigênica , Homologia de Sequência do Ácido Nucleico
20.
Cytogenet Genome Res ; 117(1-4): 213-20, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17675862

RESUMO

Previous studies in the chicken have identified a single microchromosome (GGA16) containing the ribosomal DNA (rDNA) and two genetically unlinked MHC regions, MHC-B and MHC-Y. Chicken DNA sequence from these loci was used to develop PCR primers for amplification of homologous fragments from the turkey (Meleagris gallopavo). PCR products were sequenced and overgo probes were designed to screen the CHORI 260 turkey BAC library. BAC clones corresponding to the turkey rDNA, MHC-B and MHC-Y were identified. BAC end and subclone sequencing confirmed identity and homology of the turkey BAC clones to the respective chicken loci. Based on subclone sequences, single-nucleotide polymorphisms (SNPs) segregating within the UMN/NTBF mapping population were identified and genotyped. Analysis of SNP genotypes found the B and Y to be genetically unlinked in the turkey. Silver staining of metaphase chromosomes identified a single pair of microchromosomes with nucleolar organizer regions (NORs). Physical locations of the rDNA and MHC loci were determined by fluorescence in situ hybridization (FISH) of the BAC clones to metaphase chromosomes. FISH clearly positioned the rDNA distal to the Y locus on the q-arm of the MHC chromosome and the MHC-B on the p-arm. An internal telomere array on the MHC chromosome separates the B and Y loci.


Assuntos
Cromossomos/genética , Cromossomos/imunologia , Antígenos de Histocompatibilidade/genética , Antígenos de Histocompatibilidade/imunologia , Perus/genética , Perus/imunologia , Animais , Sequência de Bases , Clonagem Molecular , Bases de Dados de Ácidos Nucleicos , Genômica , Hibridização In Situ , Metáfase , Técnicas de Amplificação de Ácido Nucleico , Mapeamento Físico do Cromossomo
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