分别提取锯缘青蟹Scylla serrata卵巢、精巢、眼柄、促雄腺的总RNA,等量混合后经oligotex试剂分离mRNA,利用SMART(Switch mechanisms at the 5′end of RNA transcript)技术及DSN(duplex-specific nuclease)的特性构建线性化cDNA质粒文...分别提取锯缘青蟹Scylla serrata卵巢、精巢、眼柄、促雄腺的总RNA,等量混合后经oligotex试剂分离mRNA,利用SMART(Switch mechanisms at the 5′end of RNA transcript)技术及DSN(duplex-specific nuclease)的特性构建线性化cDNA质粒文库。经检测文库约含有5.3×107重组子,扩增后的文库含有1011以上重组子。文库的插入cDNA长度为0.5—2.5kb,重组率大于98%。从构建的文库中随机挑取5 202个克隆进行大规模EST测序,获得3 346条ESTs,其中长度大于100bp的高质量ESTs 2 697条。经拼接与软件分析,2 697 ESTs代表了2 522个Unigenes,包括2 355个Singlets和167个Contigs,冗余率6.49%。EST序列经生物信息学分析(BLASTX,e<10-6),发现了Sry-like protein C、Sox14 protein、Sox4b、sex-determining protein fem-1、vitellogenin、vitellogenin receptor、cyclin A、cathepsin C和cyclin-dependent kinase 2等与性别分化和性腺发育相关的基因,以及热休克蛋白家族、泛素系统、抗氧化防御体系和抗病相关的功能基因。该文库具有较高的质量和利用价值,可为更大规模EST分析及采用基因芯片技术开展锯缘青蟹性腺发育及性别分化相关功能基因的研究奠定基础。展开更多
人类基因组计划的完成(2001年)宣告了后基因组时代的到来,也掀起新一轮的RNA研究热潮.作为后基因组时代的科学前沿,RNA组学近年来成为生命科学领域的研究热点,各种新型ncRNA的发现,让人们对遗传信息表达调控网络有了新的认识.结合RNA领...人类基因组计划的完成(2001年)宣告了后基因组时代的到来,也掀起新一轮的RNA研究热潮.作为后基因组时代的科学前沿,RNA组学近年来成为生命科学领域的研究热点,各种新型ncRNA的发现,让人们对遗传信息表达调控网络有了新的认识.结合RNA领域的最新研究进展,《中国科学C辑:生命科学》(Science in China Series C-Life Sciences)2009年第3期的8篇述评,从动植物小分子非编码RNA、miRNA与细胞分化发育、miRNA与肿瘤发生及诊断治疗的靶点、核酶的结构与功能、遗传印记起源、miRNA基因簇的进化等多个方面进行了综述,展现了ncRNA领域的研究现状和发展前景.展开更多
With the support by the National Natural Science Foundation of China,the collaborative team led by Prof.Miguel Esteban,Prof.Zhang BiLiang and Prof.Bao XiChen(鲍习琛)from Guangzhou Institutes of Biomedicine and Health,...With the support by the National Natural Science Foundation of China,the collaborative team led by Prof.Miguel Esteban,Prof.Zhang BiLiang and Prof.Bao XiChen(鲍习琛)from Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences have developed a new methodology(Capture of the newly transcribed RNA Interactome using ClicK chemistry;RICK)to systematically identify展开更多
The advent of high throughput technologies has revealed that mammalian genomes are pervasively transcribed, most for long noncoding RNAs （lncRNAs, at least 200 nt long）. Thousands of lncRNAs from intergenic regions ...The advent of high throughput technologies has revealed that mammalian genomes are pervasively transcribed, most for long noncoding RNAs （lncRNAs, at least 200 nt long）. Thousands of lncRNAs from intergenic regions （large in- tergenic noncoding RNA, lincRNA） have been uncovered by massive deep sequencing from the repertoire of polyadenylat- ed （poly（A）＋） RNAs, together with multiple chromatin land-scapes. These lncRNAs are messenger RNA （mRNA）-like, with linear signatures of 5＇ mVG caps and 3＇ poly（A）＋ tails. Unex- pectedly, mammalian transcriptomes are even more complex with the expression of RNAs without polyadenylated tails （poly（A）- RNAs） [1], leading to the identification of new lncRNA formats, such as circular RNAs.展开更多
基金Supported by the National Natural Science Foundation of China(31060042,31160177)Yunnan Province and Minister Key Subject,University Key Lab and Share Platform
文摘人类基因组计划的完成(2001年)宣告了后基因组时代的到来,也掀起新一轮的RNA研究热潮.作为后基因组时代的科学前沿,RNA组学近年来成为生命科学领域的研究热点,各种新型ncRNA的发现,让人们对遗传信息表达调控网络有了新的认识.结合RNA领域的最新研究进展,《中国科学C辑:生命科学》(Science in China Series C-Life Sciences)2009年第3期的8篇述评,从动植物小分子非编码RNA、miRNA与细胞分化发育、miRNA与肿瘤发生及诊断治疗的靶点、核酶的结构与功能、遗传印记起源、miRNA基因簇的进化等多个方面进行了综述,展现了ncRNA领域的研究现状和发展前景.
文摘With the support by the National Natural Science Foundation of China,the collaborative team led by Prof.Miguel Esteban,Prof.Zhang BiLiang and Prof.Bao XiChen(鲍习琛)from Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences have developed a new methodology(Capture of the newly transcribed RNA Interactome using ClicK chemistry;RICK)to systematically identify
文摘The advent of high throughput technologies has revealed that mammalian genomes are pervasively transcribed, most for long noncoding RNAs （lncRNAs, at least 200 nt long）. Thousands of lncRNAs from intergenic regions （large in- tergenic noncoding RNA, lincRNA） have been uncovered by massive deep sequencing from the repertoire of polyadenylat- ed （poly（A）＋） RNAs, together with multiple chromatin land-scapes. These lncRNAs are messenger RNA （mRNA）-like, with linear signatures of 5＇ mVG caps and 3＇ poly（A）＋ tails. Unex- pectedly, mammalian transcriptomes are even more complex with the expression of RNAs without polyadenylated tails （poly（A）- RNAs） [1], leading to the identification of new lncRNA formats, such as circular RNAs.
