This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method...
This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.
This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.
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제안 방법
The seminal plasma protein profile, using 2-DE, has not been described yet in Hanwoo. Therefore, the objective of this study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo.
이론/모형
Seminal plasma protein was extracted from the freeze dried seminal plasma powder using freeze dryer by M-PER Mammalian Protein Extraction Reagent (Thermo, USA). Proteins were refined by clean-up kit (GE Healthcare, USA), and protein content was quantified with 300 Mg using Bradford's method.
This study have analyzed seminal plasma protein from Hanwoo using the 2-DE method. The distribution of Hanwoo seminal plasma proteins on Comassie Brilliant Blue (CBB) and silver stained gels are shown in Fig.
This study was performed for a comprehensive protein profile of seminal plasma from Hanwoo (Korean Native Cattle) using the 2-DE. And, 2-DE spots were visualized through CBB and silver stain (Fig.
성능/효과
In conclusion, this study is protein profile of seminal plasma from Hanwoo using the 2-DE, and we found that 12 new proteins has not been previously described in the bovine seminal plasma study. So far, this the role of those proteins on the fertility of the male Hanwoo are still unknown.
후속연구
Therefore, we suggest that additional research are necessary to define the types of proteins affecting fertility of male and the mechanisms of their actions. Additional research may contribute to the development of strategies to improve the fertilizing ability of semen in Hanwoo.
So far, this the role of those proteins on the fertility of the male Hanwoo are still unknown. Therefore, we suggest that additional research are necessary to define the types of proteins affecting fertility of male and the mechanisms of their actions. Additional research may contribute to the development of strategies to improve the fertilizing ability of semen in Hanwoo.
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