Micro RNAs（mi RNAs） are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal.To discover novel mi RNAs a...Micro RNAs（mi RNAs） are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal.To discover novel mi RNAs and identify the differential expression of them in ovine ovary and testis tissues, the study constructed two libraries by using next-generation sequencing technologies（Solexa high-throughput sequencing technique）.As a result, 9 321 775 and 9 511 538 clean reads were obtained from the ovary and testis separately, which included 130 562（90 genes of ovary） and 56 272（85 genes of testis） of known mi RNAs and 486 potential novel mi RNAs reads.In this study, a total of 65 conserved mi RNAs were significantly differentially expressed（P〈0.01） between the two samples.Among them, 28 mi RNAs were up-regulated and 3 mi RNAs were down-regulated on ovary compared with testis.In addition, the known mi RNAs with the highest expression level（5 mi RNAs） and 30 novel mi RNAs with the functions related to reproduction were validated using the real-time quantitative RT-PCR.Moreover, the gene ontology（GO） annotation and Kyoto encyclopedia of genes and genomes（KEGG） pathway analysis showed that differentially expressed mi RNAs were involved in ovary and testis physiology, including signal transduction, gonad development, sex differentiation, gematogenesis, fertilization and embryo development.The results will be helpful to facilitate studies on the regulation of mi RNAs during ruminant reproduction.展开更多
The current research presents the protein changes in plasma from healthy dairy cows and clinical mastitic cows using two- dimensional gel electrophoresis （2-DE）. After staining with silver nitrate and Coomassie Blue...The current research presents the protein changes in plasma from healthy dairy cows and clinical mastitic cows using two- dimensional gel electrophoresis （2-DE）. After staining with silver nitrate and Coomassie Blue, differential expression proteins were detected by PDQuest 7.4 software, and then subjected to ion trap mass spectrometer equipped with a Surveyor HPLC System, differential spots of protein were identified. Three protein spots that originated from preparation gels were identified to be two proteins. Overall, haptoglobin precursor was up-regulated in cows infected with clinical mastitis and could be a mastitis-associated diagnostic marker, whereas SCGB 2A1 （secretoglobin, family 2A, member 1） was down-regulated protein. Plasma protein expression patterns were changed when cows were infected with mammary gland inflammation; it suggests that analysis of differential expression protein might be useful to clarify the mechanisms involved in the pathophysiology, and find new diagnostic markers of mastitis and potential protein targets for treatment.展开更多
This study is first to investigate proteomic changes in sheep sperm induced by carbon ion radiation using two-dimensional electrophoresis （2-DE） analysis in the project of breeding a new variety of sheep. Differenti...This study is first to investigate proteomic changes in sheep sperm induced by carbon ion radiation using two-dimensional electrophoresis （2-DE） analysis in the project of breeding a new variety of sheep. Differential expression proteins were detected using the PDQuest 8.0 software after staining with Coomassie blue. Valid spots were then analyzed through liquid chromatography tandem mass spectrometry （LC-MS/MS）. Among the 480 total protein spots displayed in 2-D gels, 6 specific protein spots were observed in sperm gels. A search against protein sequences in the National Center for Biotechnology Information databases （NCBI） indicated that differentially expressed proteins correspond to two proteins, identified to be enolase and transcription factor AP-2-alpha （TFAP-2c0. The two proteins were up-regulated in the irradiated sperm. To the best of our knowledge, this study is the first to identify proteomic changes induced by carbon ion radiation in sheep sperm. The analysis of differential expression protein may be useful in identifying new breeding markers in sheep reproduction and in clarifying the mechanisms involved in irradiation or space breeding.展开更多
基金supported by the Department of Agriculture and Animal Husbandry of Gansu,China(2011zx08008-003)the Ministry of Agriculture of China(2009ZX08008008-002)the Scientific Research Basic Business Expenses of Colleges and Universities,China(2011zx08008-003)
文摘Micro RNAs（mi RNAs） are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal.To discover novel mi RNAs and identify the differential expression of them in ovine ovary and testis tissues, the study constructed two libraries by using next-generation sequencing technologies（Solexa high-throughput sequencing technique）.As a result, 9 321 775 and 9 511 538 clean reads were obtained from the ovary and testis separately, which included 130 562（90 genes of ovary） and 56 272（85 genes of testis） of known mi RNAs and 486 potential novel mi RNAs reads.In this study, a total of 65 conserved mi RNAs were significantly differentially expressed（P〈0.01） between the two samples.Among them, 28 mi RNAs were up-regulated and 3 mi RNAs were down-regulated on ovary compared with testis.In addition, the known mi RNAs with the highest expression level（5 mi RNAs） and 30 novel mi RNAs with the functions related to reproduction were validated using the real-time quantitative RT-PCR.Moreover, the gene ontology（GO） annotation and Kyoto encyclopedia of genes and genomes（KEGG） pathway analysis showed that differentially expressed mi RNAs were involved in ovary and testis physiology, including signal transduction, gonad development, sex differentiation, gematogenesis, fertilization and embryo development.The results will be helpful to facilitate studies on the regulation of mi RNAs during ruminant reproduction.
基金supported by the China International Sci & Tech Cooperation Project (2005 DFA30720)
文摘The current research presents the protein changes in plasma from healthy dairy cows and clinical mastitic cows using two- dimensional gel electrophoresis （2-DE）. After staining with silver nitrate and Coomassie Blue, differential expression proteins were detected by PDQuest 7.4 software, and then subjected to ion trap mass spectrometer equipped with a Surveyor HPLC System, differential spots of protein were identified. Three protein spots that originated from preparation gels were identified to be two proteins. Overall, haptoglobin precursor was up-regulated in cows infected with clinical mastitis and could be a mastitis-associated diagnostic marker, whereas SCGB 2A1 （secretoglobin, family 2A, member 1） was down-regulated protein. Plasma protein expression patterns were changed when cows were infected with mammary gland inflammation; it suggests that analysis of differential expression protein might be useful to clarify the mechanisms involved in the pathophysiology, and find new diagnostic markers of mastitis and potential protein targets for treatment.
基金supported by the Key Scientific Technology Research Projects of Gans u Province, China (1102NKDA022)the Major Program of Ministry of Agriculture to Cultivate New Varieties of Genetically ModifiedOrganisms of China (2008ZX08008-003)+1 种基金the National Basic Research Program of Chinathe National Natural Science Foundation of China (2010CB834202)
文摘This study is first to investigate proteomic changes in sheep sperm induced by carbon ion radiation using two-dimensional electrophoresis （2-DE） analysis in the project of breeding a new variety of sheep. Differential expression proteins were detected using the PDQuest 8.0 software after staining with Coomassie blue. Valid spots were then analyzed through liquid chromatography tandem mass spectrometry （LC-MS/MS）. Among the 480 total protein spots displayed in 2-D gels, 6 specific protein spots were observed in sperm gels. A search against protein sequences in the National Center for Biotechnology Information databases （NCBI） indicated that differentially expressed proteins correspond to two proteins, identified to be enolase and transcription factor AP-2-alpha （TFAP-2c0. The two proteins were up-regulated in the irradiated sperm. To the best of our knowledge, this study is the first to identify proteomic changes induced by carbon ion radiation in sheep sperm. The analysis of differential expression protein may be useful in identifying new breeding markers in sheep reproduction and in clarifying the mechanisms involved in irradiation or space breeding.
