Establishment of the optimum two-dimensional electrophoresis system of ovine ovarian tissue

Abstract

ABSTRACT. 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 h→500 V, 1 h→1000 V 1 h→1000-9000 V, 6 h→80,000 VH→500 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 6529 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 13 (3): 6528-6538 (2014) Optimum ovine ovarian two-dimensional electrophoresis system sample preparation and 2-DE using tissues from other animals.

Effect of high proportion concentrate dietary on Yak jejunal structure, physiological function and protein composition during cold season

“…There were 3 replicates in each group. Jejunal proteins were extracted by the lysis buffer method 12 . Digestion of the protein (250 μg for each sample) was performed according to the FASP procedure.…”

Effect of high proportion concentrate dietary on Yak jejunal structure, physiological function and protein composition during cold season

“…There were 3 replicates each group. Jejunum proteins were extracted by lysis buffer method (7M Urea, 2M Thiourea, 4% CHAPS(w/v), 100 mM DTT, 60 mM Tris-HCl, 0.5% pH 3-10 Ampholine, 20μL proteinase inhibiors) [11] . Digestion of the protein (250 μg for each sample) was performed according to the FASP procedure described.…”

Eukaryotic expression, Co-IP and MS identify BMPR-1B protein–protein interaction network

“…Consequently, upon binding with BMP ligands, BMPR-IIB transphosphorylates the GS domain of the BMPR-1B, which leads to the activation of downstream cascades and the inactivation of the partial receptor [8]. As a result of this, the inhibiting effect of steroidogenesis on GDF5 and BMP4 is weakened, the SMAD state of expression and phosphorylation is changed, the synthesis of estradiol by FSH induction is promoted, the synthesis and secretion of progesterone is inhibited, granular cell differentiation of mutation ewes and follicular maturation are accelerated, and ovulation is increased [9]. Although little is known about the follicular regulatory mechanism of BMPR-1B in single, twins, and multiple prolific ewes, these properties of BMPR-1B can provide new concepts for studies of ewe follicular development and litter size.…”