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
以小反刍兽疫病毒(peste des petits ruminants virus,PPRV)疫苗株Nigeria75/1基因序列为基础,构建了融合表达PPRV囊膜蛋白F和H胞外区的真核重组质粒pcDNA3.1-F-H,在悬浮CHO细胞中进行分泌表达。离心收集转染后的细胞培养上清进行亲和...以小反刍兽疫病毒(peste des petits ruminants virus,PPRV)疫苗株Nigeria75/1基因序列为基础,构建了融合表达PPRV囊膜蛋白F和H胞外区的真核重组质粒pcDNA3.1-F-H,在悬浮CHO细胞中进行分泌表达。离心收集转染后的细胞培养上清进行亲和层析纯化,纯化样品经Western-blot鉴定,融合蛋白与PPRV阳性血清和His标签抗体都能发生免疫反应。使用纯化后的融合蛋白免疫BALB/c小鼠,可诱导小鼠体内产生针对PPRV的特异性抗体。上述研究结果为小反刍兽疫新型亚单位疫苗的研制奠定了基础。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.
文摘以小反刍兽疫病毒(peste des petits ruminants virus,PPRV)疫苗株Nigeria75/1基因序列为基础,构建了融合表达PPRV囊膜蛋白F和H胞外区的真核重组质粒pcDNA3.1-F-H,在悬浮CHO细胞中进行分泌表达。离心收集转染后的细胞培养上清进行亲和层析纯化,纯化样品经Western-blot鉴定,融合蛋白与PPRV阳性血清和His标签抗体都能发生免疫反应。使用纯化后的融合蛋白免疫BALB/c小鼠,可诱导小鼠体内产生针对PPRV的特异性抗体。上述研究结果为小反刍兽疫新型亚单位疫苗的研制奠定了基础。
