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Polimorfismo no Gene que Codifica a ?-lactoglobulina e associa??o com caracter?sticas de produ??o em caprinos leiteiros / Polymorphism in the gene encoding the ?-lactoglobulin and Association with Traditional Production in Dairy GoatsRego, Ramon de Sousa 26 August 2014 (has links)
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Previous issue date: 2014-08-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / The protein quality of milk of goats and digestibility of the lipid fraction are important factors that stand out when compared to milk from cows. The ?-lactoglobulin is the higher abundant protein in whey ruminants being produced in the mammary gland during lactation. It may represent up to 12% of the total protein. We seek to present work was to evaluate the gene ?-Lactoglobulin (BLG) with their genetic polymorphisms in the promoter regions 5 'and 3' UTR and associate them the characteristics of milk production in experimental goat herd. For this 150 goats (Capra hircus) of Saanen and Alpine, genotyped for polymorphisms of the BLG gene were used. For genotyping 10 mL of blood per animal were collected. The blood was used for DNA extraction and amplification of two fragments of the BLG gene by means of polymerase chain reaction (PCR). The amplified fragments were submitted to electrophoresis on polyacrylamide gel and evaluated by by restriction fragment length polymorphism (PCR-RFLP). The SmaI and SacII enzymes were used for the promoter (promoter + exon 1 region) region and the region of exon 7 (exon 7 + region 3 '), respectively. The differences in cutting patterns were visualized on a polyacrylamide gel. In the promoter region of the single nucleotide polymorphism (SNP) at position -60 (C/T) was identified and was associated with the percentage of protein in milk. This result suggests a relationship between genotypes of the promoter region of the BLG gene and the protein level of the milk. The presence of two polymorphisms +4641 (I2/I3) and +4601 (A/G) was observed in the region of exon 7. The I2-I3 variation is characterized by a repeat sequence of 10 bp, varying in two and three times, and it being possible to identify by electrophoresis. The I3 variation was often very low, and there was no association between this polymorphism and any production trait. Polymorphism +4601 (G/A) was identified by enzyme SacII digestion. Alleles were identified frequencies close to those reported by other authors and were associated with the percentage of fat in milk goats, and the animals with the S1S2 genotype had higher fat percentage than S1S1 animals and S2S2. The increased production of lipid content may be related to the characteristic of transporting fatty acids ?-lactoglob / A qualidade proteica do leite de cabras e a digestibilidade da fra??o lip?dica s?o importantes fatores que o destacam, quando comparado ao leite de vacas. A ?-Lactoglobulina ? a prote?na de maior abundancia no soro do leite em ruminantes sendo produzida na gl?ndula mam?ria durante o per?odo de lacta??o. Ela pode representar at? 12% do total proteico. Buscamos com o presente trabalho, avaliar o gene da ?-Lactoglobulina (BLG) com seus polimorfismos gen?ticos nas regi?es promotoras 5? e 3? UTR e associ?-los as caracter?sticas de produ??o de leite no rebanho caprino experimental. Para isso foram utilizadas 150 cabras (Capra hircus) das ra?as Saanen e Alpinas, genotipadas para os polimorfismos do gene BLG. Para a genotipagem foram coletados 10 mL de sangue por animal. O sangue foi utilizado para a extra??o de DNA e amplifica??o de 2 fragmentos do gene BLG por interm?dio da rea??o em cadeia da polimerase (PCR). Os fragmentos amplificados foram ent?o submetidos ? eletroforese em gel de poliacrilamida e avaliados pelo polimorfismo no tamanho do fragmento por restri??o (PCR-RFLP). Foram utilizadas as enzimas SmaI e SacII, para a regi?o promotora (regi?o promotora + ?xon 1) e para a regi?o do ?xon 7 (?xon 7 + regi?o 3?), respectivamente. As diferen?as nos padr?es de corte foram visualizadas em gel de poliacrilamida. Na regi?o promotora o polimorfismo de base ?nica (SNP) na posi??o -60 (C/T) foi identificado e apresentou associa??o com a percentagem de prote?nas no leite. Esse resultado sugere uma rela??o entre os gen?tipos da regi?o promotora do gene BLG e o n?vel proteico do leite. Foi observado na regi?o do ?xon 7 a presen?a de dois polimorfismos +4641 (I2/I3) e +4601 (A/G). A varia??o I2-I3 ? caracterizada pela repeti??o de uma sequ?ncia de 10 pb, variando em duas e tr?s vezes, e sendo poss?vel identifica-la por eletroforese. A varia??o I3 teve frequ?ncia muito baixa, e n?o houve associa??o entre esse polimorfismo e nenhuma caracter?stica de produ??o. O polimorfismo +4601 (A/G) foi identificado por meio da digest?o da enzima SacII. Os alelos identificados tiveram frequ?ncias pr?ximas ?s relatadas por outros autores e apresentaram associa??o com a percentagem de gordura no leite cabras, sendo que os animais com o gen?tipo S1S2 apresentaram maior percentagem de gordura que animais S1S1 e S2S2. A maior produ??o de conte?do lip?dico pode estar relacionada com a caracter?stica de transporte de ?cidos graxos da ?-Lactoglobulina
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