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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2019.tde-20191218-131659
Document
Author
Full name
Luciana Correia de Almeida Regitano
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1991
Supervisor
Title in Portuguese
Polimorfismo de proteínas e de DNA em eqüinos (Equus caballus)
Keywords in Portuguese
DNA
EQUINOS
MARCADOR GENÉTICO
POLIMORFISMO
PROTEÍNAS
Abstract in Portuguese
Com o objetivo de avaliar o polimorfismo de alguns marcadores genéticos detectáveis no sangue de eqüino (Equus caballus) foram estudados, no presente trabalho, os sistemas: albumina e esterase do soro; 6-fosfogluconato-desidrogenaseesterase (PGD) e fosfohexose-isomerase (PHI) dos eritrócitos; e o padrão de restrição do DNA (RFLP) para sondas de seqüências ao acaso de DNA genômico de eqüinos. A amostra foi constituída de 9 famílias completas e 4 famílias incompletas (mãe e progênie) de eqüinos Mangalarga Marchador, provenientes de uma criação localizada em São Sebastião do Paraíso (MG), totalizando 27 animais. Uma família de pôneis da raça Shetland foi incluída nas análises de DNA. A determinação dos fenótipos de albumina e esterase foi feita por eletroforese em gel de amido a pH 5,4. Três fenótipos de albumina foram observados, eles controlados por dois alelos codominantes, AIA e AlB, com freqüências de 0,54 e 0,46, respectivamente para este estudo. O locus Es da esterase também apresentou dois alelos codominantes, F e I, com freqüências de 0,42 e 0,58, respectivamente. Apesar do reduzido tamanho da amostra, 12 animais na geração parental, a população se encontrou em equilíbrio de Hardy-Weinberg para esses dois loci. Os dois sistemas eritrocitários estudados, PGD e PHI, mostraram-se monomórficos em eletroforese em gel de amido a pH 7,2. Para o estudo de RFLP foram utilizadas sondas constituídas de fragmentos ao acaso de DNA de eqüino, digerido com a enzima Sau3AI. Essas sondas foram isoladas de banco de DNA genômico construído no vetor pUC19. Três métodos de hibridização e três membranas de nylon foram testados com resultados significativamente diferentes na qualidade das hibridizações obtidas. Uma das sondas testadas, a sonda pESa, detectou a ocorrência de dois RFLPs de 1,9 kb e 1,3 kb para a enzima HindIII, e dois RFLPs (4 kb e 3,2 kb) para a enzima EcoRI, demonstrando a utilidade das sondas de seqüências ao acaso na obtenção de novos marcadores genéticos.
Title in English
Protein and DNA polymorphism in equines (Equus caballus)
Abstract in English
ln order to evaluate the polymorphism of some blood genetic markers in equine (Equus caballus), the following systems were studied: serum albumin, serum esterase, red cell 6-phosphogluconate dehydrogenase (PGD) and red cell phosphohexose isomerase (PHI) and the restriction fragment lenght poplymorphism (RFLP) for random equine DNA sequence probes. The sample was composed of 9 complete and 4 incomplete (mother and offspring) horse families of the Mangalarga Marchador breed, from one farm in São Sebastião do Paraíso (Minas Gerais State), in a total of 27 animals. One family of the Shetland pony breed was included in the DNA analysis. The albumin and esterase phenotypes were determined by starch gel electrophoresis at pH 5.4. Three albumin phenotypes two codominant alleles, AlA and AlB, with frequencies of 0.54 and 0.46, respectively for this study. Two codominant alleles were also detected in the esterase locus Es, alleles F and I, with frequencies of 0.42 and 0.58, respectively. Despite the small sample size, only 12 animals in the parental generation, the population was in Hardy-Weinberg equilibrium for these two loci. The two red cell systems studied, PGD and PHI, were determined by starch gel electrophoresis at pH 7.2, and showed to be monomorphic in the sample. For the RFLP study, random sequences of equine DNA Sau3AI digests were used as probes. These probes were isolated from genomic DNA lib:rary cloned in the vector pUC19. Three hybridization methods and three nylon membranes were tested. Significant differences in the resulting quality of hybridizations were found among them. Two RFLPs of 1.9 kb and 1.3 kb for HindIII digests, and two others (4 kb and 3.2 kb) for EcoRI digests were detected by one of the probes, pES3.These results indicate the usefulness of random sequence probes in the detection of new genetic markers.
 
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Publishing Date
2019-12-19
 
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