RESUMO
Lambing performance of sheep is the most important economic trait and is regarded as a critic factoring affecting the productivity in sheep industry. Ovary plays the most roles in lambing trait. To establish the optimum two-dimensional electrophoresis system (2-DE) of ovine ovarian tissue, the common protein extraction methods of animal tissue (trichloroacetic acid/acetone precipitation and direct schizolysis methods) were used to extract ovine ovarian protein, and 17-cm nonlinear immobilized PH 3-10 gradient strips were used for 2-DE. The sample handling, loading quantity of the protein sample, and isoelectric focusing (IEF) steps were manipulated and optimized in this study. The results indicate that the direct schizolysis III method, a 200-µg loading quantity of the protein sample, and IEF steps II (20°C active hydration, 14 hâ500 V, 1 hâ1000 V 1 hâ1000-9000 V, 6 hâ80,000 VHâ500 V 24 h) are optimal for 2-DE analysis of ovine ovarian tissue. Therefore, ovine ovarian tissue proteomics 2-DE was preliminarily established by the optimized conditions in this study; meanwhile, the conditions identified herein could provide a reference for ovarian sample preparation and 2-DE using tissues from other animals.
Assuntos
Eletroforese em Gel Bidimensional/métodos , Ovário/metabolismo , Proteoma/análise , Proteômica/métodos , Acetona/química , Animais , Precipitação Química , Feminino , Concentração de Íons de Hidrogênio , Focalização Isoelétrica , Proteoma/isolamento & purificação , Reprodutibilidade dos Testes , Ovinos , Ácido Tricloroacético/químicaRESUMO
The growth hormone gene plays an important role in the physiological function of an organism. The current study aimed to investigate the correlation between polymorphisms in the 5' regulatory region, exon 4, and 3' untranslated region (UTR) of the sheep GH gene and sheep growth traits. The DNA from 510 adult sheep was analyzed by DNA sequencing and polymerase chain reaction single-strand conformation polymorphism. Two alleles (A and B) and 3 genotypes (AA, AB, and BB), 2 alleles (A and B) and 3 genotypes (AA, AB, and BB), and 3 alleles (A, B, and C) and 4 genotypes (AA, AB, BB, and AC) were found within the 5' regulatory region, exon 4, and 3' UTR, respectively. In Tibetan sheep, the association analysis indicated that there were statistically significant differences in the scores of weight, length, and heart girth within the 5' regulatory region; weight, length, wither height, and heart girth within exon 4; and weight, length, wither height, and heart girth within the 3' UTR among the different genotypes. For exon 4, Poll Dorset sheep individuals with genotype AA showed a lower score than those of genotypes BB and AB (P<0.05). With regard to the 3' UTR, Poll Dorset sheep with genotype AC showed higher scores than those of genotypes AA and AB (P<0.05).