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用PCR法构建乳腺特异性表达非乳蛋白基因的研究 被引量:1
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作者 郭继彤 李雪玲 +2 位作者 于海泉 王达珍 张肇英 《内蒙古大学学报(自然科学版)》 CAS CSCD 1997年第6期830-835,共6页
将PCR法分别从绵羊和人血基因组DNA中扩增出的绵羊β-乳球蛋白基因(BLG)5′端调控区898bp的片段和人骨髓单核细胞表面分化抗原CD14基因(hCD14)成熟肽1245bp的片段连接.连接片段插入pUC19Ec... 将PCR法分别从绵羊和人血基因组DNA中扩增出的绵羊β-乳球蛋白基因(BLG)5′端调控区898bp的片段和人骨髓单核细胞表面分化抗原CD14基因(hCD14)成熟肽1245bp的片段连接.连接片段插入pUC19EcoRI-KpnI位点,构建成pBLG-hCD14质粒.该质粒经KpnI及EcoRI-HindII酶切、PCR法扩增BLG和hCD14分析后,进一步用32P-BLG探针杂交确证.用EcoRI-HindII酶切质粒片段BLG-hCD14产生转基因小鼠以检测BLG-hCD14在乳腺的表达能力.用免疫杂交法证实在转基因阳性小鼠乳汁中表达了hCD14. 展开更多
关键词 PCR法 基因动物 非乳蛋白基因 基因表达
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Evolutionary annotation of conserved long non-coding RNAs in major mammalian species 被引量:3
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作者 BU DeChao LUO HaiTao +4 位作者 JIAO Fei FANG ShuangSang TAN ChengFu LIU ZhiYong ZHAO Yi 《Science China(Life Sciences)》 SCIE CAS CSCD 2015年第8期787-798,共12页
Mammalian genomes contain tens of thousands of long non-coding RNAs(lnc RNAs) that have been implicated in diverse biological processes. However, the lnc RNA transcriptomes of most mammalian species have not been esta... Mammalian genomes contain tens of thousands of long non-coding RNAs(lnc RNAs) that have been implicated in diverse biological processes. However, the lnc RNA transcriptomes of most mammalian species have not been established, limiting the evolutionary annotation of these novel transcripts. Based on RNA sequencing data from six tissues of nine species, we built comprehensive lnc RNA catalogs(4,142–42,558 lnc RNAs) covering the major mammalian species. Compared to protein-coding RNAs, expression of lnc RNAs exhibits striking lineage specificity. Notably, although 30%–99% human lnc RNAs are conserved across different species on DNA locus level, only 20%–27% of these conserved lnc RNA loci are detected to transcription, which represents a stark contrast to the proportion of conserved protein-coding genes(48%–80%). This finding provides a valuable resource for experimental scientists to study the mechanisms of lnc RNAs. Moreover, we constructed lnc RNA expression phylogenetic trees across nine mammals and demonstrated that lnc RNA expression profiles can reliably determine phylogenic placement in a manner similar to their coding counterparts. Our data also reveal that the evolutionary rate of lnc RNA expression varies among tissues and is significantly higher than those for protein-coding genes. To streamline the processes of browsing lnc RNAs and detecting their evolutionary statuses, we integrate all the data produced in this study into a database named Phylo NONCODE(http://www.bioinfo.org/phylo Noncode). Our work starts to place mammalian lnc RNAs in an evolutionary context and represent a rich resource for comparative and functional analyses of this critical layer of genome. 展开更多
关键词 IncRNA CONSERVATION evolution
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