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Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes

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cell Research 2003年第4期13卷 251-263页

作者： YING CHEN, ZHI Xu HE, AILIAN LIU, KAI WANG, WEN WEI MAO, JIAN XIN CHU, YONG LU, ZHENG Fu FANG, YING TANG SHI, QING ZHANG YANG, DA YUAN CHEN, MIN RANG WANG, JIN SONG LI, SHAO LIANG HUANG, XIANG YIN KONG, YAO ZHOU SHI, Zm QIANG WANG, JIA HuI XIA, ZHI GAO LONG, ZHI GANG XUE, WEN XIANG DING, HuI ZHEN SHENG Center for Developmental biology, shanghai Second Medical University, shanghai 200092, china laboratory of Stem cell biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 1665 Kong Jiang Road, shanghai 200092, china Center for Stem cell Research, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510120,china state key laboratory of Reproductive biology, institute of Zoology, chinese academy of sciences, 19 Zhong-guan Cun Road, Haidian District, Beijing 100080, china Health Science Center, shanghai institutes of biological sciences, chinese academy of sciences and shanghai Second Medical University, shanghai 200025, china National laboratory of Medical Genetics, Central South University, 88 Xiangya Road, Changsha 410078, china Xinhua Hospital, shanghai Second Medical University, 1665 Kong Jiang Road, shanghai 200092, china 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 3Center for Stem cell Research the Second Affiliated Hospital Sun Yat-Sen University Guangzhou 510120 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2laboratory of Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 4state key laboratory of Reproductive biology Institute of Zoology Chinese Academy of Sciences 19 Zhong-guan Cun Road Haidian District Beijing 10008

To solve the problem of immune incompatibility, nuclear transplantation has been envisaged as a means to produce cells or tissues for human autologous transplantation. Here we have derived embryonic stem cells by the transfer of human somatic nuclei into rabbit oocytes. The number of blastocysts that developed from the fused nuclear transfer was comparable among nuclear donors at ages of 5, 42, 52 and 60 years, and nuclear transfer (NT) embryonic stem cells (ntES cells) were subsequently derived from each of the four age groups. These results suggest that human somatic nuclei can form ntES cells independent of the age of the donor. The derived ntES cells are human based on karyotype, isogenicity, in situ hybridization, PCR and immunocytochemistry with probes that distinguish between the various species. The ntES cells maintain the capability of sustained growth in an undifferentiated state, and form embryoid bodies, which, on further induction, give rise to cell types such as neuron and muscle, as well

关键词： nuclear transfer (NT) somatic cell nuclear transfer (SCNT) embryonic stem cells (ES cell) therapeutic cloning rabbit oocyte.

一种黑豆蚜储存蛋白质的分离纯化及其亚基的性质

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生物化学与生物物理学报 2003年第2期35卷 189-192页

作者：甘明丁德诚周元聪中国科学院上海生命科学研究院植物生理生态研究所中国科学院上海生命科学研究院生物化学与细胞生物学研究所蛋白质组学重点实验室上海200032 上海200031

从黑豆蚜 (AphiscraccivoraKoch)血淋巴中经DE5 2纤维素离子交换层析、SephadexG 15 0凝胶过滤以及在FPLC上QSepharose分离出一种蛋白质。该蛋白质经SDS PAGE测定在非还原和还原条件下分子量均为 6 0kD左右 ,等电聚焦电泳测得其等电点约... 详细信息

从黑豆蚜 (AphiscraccivoraKoch)血淋巴中经DE5 2纤维素离子交换层析、SephadexG 15 0凝胶过滤以及在FPLC上QSepharose分离出一种蛋白质。该蛋白质经SDS PAGE测定在非还原和还原条件下分子量均为 6 0kD左右 ,等电聚焦电泳测得其等电点约为 5 .0 ,为糖蛋白。若蚜期间该蛋白质在蚜虫体内积累 ,羽化至成蚜时含量明显减少 ,为蚜虫提供发育所需氨基酸 ,在成蚜期间仍以低浓度存在。根据其分子量和氨基酸组成与含量变化动态应属不完全变态昆虫的一种持续性储存蛋白质。

关键词：黑豆蚜储存蛋白质分离纯化

水稻齿叶矮缩病毒S8基因编码的结构蛋白质及其自聚集性质

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生物化学与生物物理学报 2002年第5期34卷 565-570页

作者：吕慧涓邵朝纲龚祖埙中国科学院上海生命科学研究院生物化学与细胞生物学研究所、蛋白质组学重点实验室中国科学院上海生命科学研究院生物化学与细胞生物学研究所、蛋白质组学重点实验室上海200031

水稻齿叶矮缩病毒是一种以水稻为侵害寄主的植物呼肠孤病毒 ,其基因组 1 0条dsRNA编码的蛋白质产物的功能除S9外的大部分还未阐明。报道了该病毒菲律宾分离株S8的开放阅读框序列 ,在大肠杆菌中表达和纯化出了其 6 7kD的蛋白质产物 ,并... 详细信息

水稻齿叶矮缩病毒是一种以水稻为侵害寄主的植物呼肠孤病毒 ,其基因组 1 0条dsRNA编码的蛋白质产物的功能除S9外的大部分还未阐明。报道了该病毒菲律宾分离株S8的开放阅读框序列 ,在大肠杆菌中表达和纯化出了其 6 7kD的蛋白质产物 ,并进一步研究了该产物的功能。实验结果表明 ,S8的蛋白质产物是病毒的一种主要结构蛋白质 ,在体外具有自剪切活性和自聚集性质 ,并推测可能是病毒的内层衣壳蛋白

关键词： RRSV P8 结构蛋白质自剪切自聚集

pH-Dependent Stability of EGX, A Multi-functional cellulase from Mollusca, Ampullaria crossean

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 603-608页

作者： Wen-Ying LI2, Ji WANG1, Yan-Hong LI1, Ming DING1, Gen-Jun XU1,3, Lan-Ying LIU2, and Fu-Kun ZHAO1,3* 1key laboratory of Proteomics, institute of biochemistry and cell biology, shanghai institutes for biological sciences, the chinese academy of sciences,Graduate School of the chinese academy of sciences, shanghai 200031, china 2College of Life Science, Jilin University, Changchun 130023, china College of Life Science, Zhejiang University of Science and Technology, Hangzhou 310018, china key laboratory ofProteomics Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological Sciencesthe Chinese Academy of SciencesGraduate School of the Chinese Academy of SciencesShanghai 200031China

The cellulase activity and stability of EGX, a multi-functional cellulase previously purified from the mollusca Ampullaria crossean, was systematically studied under different pH. The pH induced con- formation and stability change of EGX have been investigated by using the intrinsic fluorescence, ANS fluorescence and CD spectrum. It has been found that the conformation and activity of this cellulase were strongly dependent on the pH. EGX was stable for both the enzyme activity and the conformation from pH 5.6 to pH 7.4. As shown by intrinsic and ANS fluorescence, no red shift of emission maximum occurred and a negligible intensity change was observed at pH 5.6–7.4. The activity of EGX remained about 80% in pH 5.6–7.4 and obviously decreased out of side the pH range. Urea-induced changes in EGX at pH 5.4 and pH 8.0 were measured by intrinsic fluorescence and CD spectrum. At pH 5.4, a significantly red shift of emission maximum occurred when the concentration of urea was 5 M compared to the concentration was 3 M at pH 8.0. The α-helix at pH 5.4 was 40.51% in the absence of urea and 31.04% in the presence of 4 M urea. At pH 8.0 the α-helix was 7.23% in the presence of 4 M urea. The data indicated that EGX was much susceptible to urea-induced unfolding at pH 8.0 and much stable at pH 5.4. The greater pH dependent stability of EGX may allow the enzyme to adequately catalyze the hydrolysis of cellulosic materials under natural or industrial extreme conditions.

关键词： multi-functional cellulase stability Ampullaria crossean fluorescence spectroscopy pH-dependent structural change CD spectrum

Two closely linked rice U14 boxC/D snoRNAs

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Progress in Natural Science:Materials International 2002年第1期 22-25页

作者： JIANG Ge, XU Feng, JIN Youxin and WANG Debao(state key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai institutes of Life sciences, chinese academy of sciences, shanghai 200031, china) state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes of Life Sciences Chinese Academy of Sciences Shanghai 200031 China

By analysis of the conserved elements in yeast U14 boxC/D snoRNA, the conserved elements in rice U14 boxC/D snoRNA have been speculated. Through computer search of the international rice genome database, two rice U14 snoRNA gene candidates are obtained. These two putative U14 snoRNA genes are closely linked on rice chromosome 2. The coding sequences of these two snoR-NAs exhibit the hallmark structure of boxC/D antisense snoRNA. They both have conserved boxC and boxD sequences and a 14nt-long complement to the sequence between 414nt and 427nt of rice 18S rRNA (according to GenBank accession no. X00755). The experimental evidence shows that these two snoRNAs are involved in the methylation of the complementary sequence of rice 18S rRNA. The existence and localization of these two snoRNAs are proved by RT-PCR and Northern blot. Further analysis shows that both of the newly found rice snoRNAs have high homology with maize U14 snoRNA, and they are named rice U14.1 snoRNA and U14.2 snoRNA respectively. The g

关键词： rice U14 BoxC/D snoRNA rRNA methylation.

A novel snoRNA can direct site-specific 2’-O-ribose methylation of snRNAs in Oryza Sativa

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A novel snoRNA can direct site-specific 2’-O-ribose methyla...

一种提高固定pH梯度(IPG)凝胶双向电泳的重复性、分辨率和通量的方法(英文)

第三届全国暨2003年中外RNA研讨会

作者： Wei Li Ge Jiang Bing Huang Youxin Jinstate key laboratory of molecular biology institute of biochemistry and cell biology shanghai institutes for biological sciences chinese academy of sciences shanghai 200031 china

＜正＞ Small nucleolar RNAs （snoRNA） are a kind of noncoding RNAs, and the vast majority of snoRNAs are involved in site-specific modifications of rRNAs. A novel box C/D snoRNA called snoR124 was found in oryza sativa, and it can direct 2’-O-ribose methylation of spliceosomal small nuclear RNAs （snRNAs）. The snoRNA has two antisense elements, and the results of primer extensions at different dNTP concentrations provide evidence that snoR124 guide 2’-O-methylations of C93 in the U4 snRNA and the T87 residue in the U5 snRNA. In addition, this snoRNA is located in a snoRNA gene cluster with another 7 snoRNAs which are identified to direct ribose methylations in rRNAs. This is consistent with the opinion that the snoRNA gene organization in plant is mainly gene cluster. The snoR124 is the first example of a snoRNA that directs modifications of RNAs other than rRNAs in plant;it will avail to get more insights into the function of snoRNAs in plant.

关键词： 2’-O-ribose methylation box C/D snoRNA snRNA oryza sativa plant

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生物化学与生物物理学报 2003年第7期35卷 611-618页

作者：袁泉安杰刘定干赵辅昆中国科学院上海生命科学研究院生物化学与细胞生物学研究所蛋白质组学重点实验室中国科学院上海生命科学研究院生物化学与细胞生物学研究所蛋白质组学重点实验室上海200031

在蛋白质组学研究中 ,双向聚丙烯酰胺凝胶电泳是现行蛋白质分离的最重要的方法之一。实验发展了一种提高固定pH梯度 (IPG)凝胶双向电泳的重复性、分辨率和通量的方法 :在一块SDS 聚丙烯酰胺凝胶上同时进行多块固定pH梯度(IPG)凝胶 (Mult... 详细信息

在蛋白质组学研究中 ,双向聚丙烯酰胺凝胶电泳是现行蛋白质分离的最重要的方法之一。实验发展了一种提高固定pH梯度 (IPG)凝胶双向电泳的重复性、分辨率和通量的方法 :在一块SDS 聚丙烯酰胺凝胶上同时进行多块固定pH梯度(IPG)凝胶 (Multi stripsononeSDSgel,MSOG)电泳。用此方法比较了人肝癌细胞、不同生长状态的人肝癌细胞、3T3细胞的蛋白质以及同一个样品在不同大小的第二向凝胶系统 (大型和中型凝胶 )的双向电泳图谱。结果表明 ,同一样品在 13cmIPGStrip双向电泳可分离 2 0 0 0以上蛋白质点且图谱蛋白质点的匹配率可超过 95 %以上。同时又可以最大程度地降低凝胶背景对蛋白质点比较分析的干扰 ,从而提高了双向电泳分离蛋白质的分辨率和通量。这些优点都有助于差异蛋白质组学特别是细胞器差异蛋白质组学研究的自动化。

关键词：双向电泳同时进行多块固定pH梯度(IPG)凝胶的方法重复性高通量蛋白质组学

FGF13A represses transcription of rRNA genes as a nucleolar protein

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FGF13A represses transcription of rRNA genes as a nucleolar ...

In Vivo Mediator Requirement for a Basal Level of Transcription

中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Liu YANG~1,Lan BAO~2,Xu ZHANG~(1,*) 1 institute of Neuroscience and state key laboratory of Neuroscience 2 institute of biochemistry and cell biology and laboratory of molecular cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai,200031,china

Objective Fibroblast growth factors(FGFs) are among the first-indentified families of growth *** are polypeptides with diverse biological activities including cell proliferation,differentiation,migration and so *** their functions are much more evident in the nervous system than in *** FGFs have N-terminal signal peptides and can be secreted ***,FGF11 subfamily(FGFs 11-14) is a special *** FGFs lack signal peptides, remain intracellularly and function within cells in a receptor-independent manner,hereafter referred to as *** growth factor 13(FGF13) is a member of the intracellular FGF homologous factor subfamily(iFGFs) and highly expressed in nervous system,however its functions are still *** The subcellular localization of FGF13A was investigated by *** interaction between FGF13A and UBF1 was investigated by coimmunoprecipitation (co-IP) *** function of FGF13A on the neurite outgrowth of DRG neurons was investigated by electroporation and ***(1) The expression of FGF13A-EGFP in transfected ND7-23 cells or dissociated primary DRG neurons was highly enriched in the nucleolus of the two types of cells and colocalized with the nucleolus marker,fibrillarin and nucleolin.(2) Overexpression of FGF13A repressed rRNA transcription through interacting with upstream binding factor 1(UBF1),a RNA pol I transcription factor.(3) Overexpression of FGF13A significantly inhibited neurite outgrowm of dissociated DRG *** Nucleolus-localized FGF13A represses rRNA transcription and therefore inhibits DRG neurite outgrowth.

关键词： iFGF UBF1 ribosomal RNA transcription neurite outgrowth

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In Vivo Mediator Requirement for a Basal Level of Transcript...