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在Yunnanese（Aγδβ）^0—地贫3′缺失端点下游鉴别到两个增强子样顺序

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生物化学与生物物理进展 2002年第1期29卷 114-119页

作者：黄小东张俊武中国医学科学院中国协和医科大学基础医学研究所

利用荧光酶报告基因系统搜索了Yunnanese (Aγδβ) 0 地中海贫血缺失 3′端点下游 11 5kb区域内的调控顺序 .确定缺失 3′端点立即下游区 1 7kb片段 ,在人红白血病细胞K5 6 2及鼠红白血病细胞MELGM979中 ,可使γ 珠蛋白基因启动子驱... 详细信息

利用荧光酶报告基因系统搜索了Yunnanese (Aγδβ) 0 地中海贫血缺失 3′端点下游 11 5kb区域内的调控顺序 .确定缺失 3′端点立即下游区 1 7kb片段 ,在人红白血病细胞K5 6 2及鼠红白血病细胞MELGM979中 ,可使γ 珠蛋白基因启动子驱动的荧光酶基因表达增加 3 8～ 4 0倍 ,而在HeLa细胞中仅增加 1 5倍 .位于缺失 3′端点约 10kb的一个长 1 4kb片段在K5 6 2和MELGM979中 ,可使γ 基因启动子驱动的荧光酶基因表达增加 2 4～2 9倍 ,而在HeLa细胞中无增加 .结果说明这两段顺序均有增强子活性 ,并且这种活性具有一定的红细胞特异性 .进一步证明 1 7kb片段内包含多个转录调节蛋白结合模体的 430bp片段包含了 1 7kb片段的大部分增强子活性 .这些结果为缺失导致增强子样顺序并入到接近Gγ 基因 ,是Yunnanese (Aγδβ) 0 地贫缺失突变体中胎儿Gγ 珠蛋白基因 ,在成人期持续活跃表达原因的假设提供了实验证据 .

关键词： Gγ-珠蛋白基因 γ(Aδβ)^0-地中海贫血增强子样顺序成人胎儿

一个可能与T细胞发育相关的新的C2H2型锌指蛋白基因的克隆(英文)

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生物化学与生物物理进展 2004年第7期31卷 600-605页

作者：杜占文张昕张俊武中国医学科学院中国协和医科大学基础医学研究所

一些在组织和细胞分化中起重要作用的蛋白质包含锌指结构域 .为了克隆分离和研究与造血细胞分化和发育成熟相关的蛋白基因 ,利用编码C2H2型锌指蛋白结构域中部分保守氨基酸序列设计简并引物 ,以骨髓cDNA为模板 ,进行PCR扩增 ,得到若干... 详细信息

一些在组织和细胞分化中起重要作用的蛋白质包含锌指结构域 .为了克隆分离和研究与造血细胞分化和发育成熟相关的蛋白基因 ,利用编码C2H2型锌指蛋白结构域中部分保守氨基酸序列设计简并引物 ,以骨髓cDNA为模板 ,进行PCR扩增 ,得到若干新的锌指蛋白基因EST .用其中一条为探针筛选人骨髓cDNA文库 ,获得了一个新的锌指蛋白基因全长cDNA ,GenBank收录号为AF2 4 6 12 6 ,长 3888bp ,包括一个完整阅读框 ,编码 6 86个氨基酸 ,包括 17个典型的和 2个非典型的C2H2模体 ,命名为HZF2 .RNA印迹、人多组织mRNA斑点杂交分析结果显示 ,其在T淋巴细胞发育和定居的器官组织胸腺、淋巴结中有较高表达 ,在脾脏、胎肝有中度表达 ,在B淋巴细胞发育的骨髓中表达很低 ,在外周血几个淋巴细胞系中仅有极微量的表达 ,提示HZF2可能对于T淋巴细胞发育和增殖有重要功能 .该基因也在脑组织的若干部位、胎盘及肾上腺有较高表达 ,在多种其他组织细胞有微量表达 ,说明其可能对维持这些组织细胞的生理功能也起一定作用 .将编码HZF2读框的DNA顺序克隆到pEGFP N1载体中 ,转染 3T3细胞 ,证明表达的HZF2 GFP融合蛋白定位于细胞核 ,这与根据HZF2蛋白结构推测其可能作为DNA结合蛋白行使调节基因转录的功能是一致的 .

关键词：锌指蛋白基因克隆 cDNA文库筛选表达谱 T细胞发育

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一种抑制pGEX载体系统本底表达的方法

生物化学与生物物理进展 1998年第1期25卷 87-89页

作者：黄蓬缪时英王琳芳中国医学科学院基础医学研究所中国协和医科大学基础

用ｐＧＥＸ载体系统体外构建了一种人精子膜蛋白片段（ＨＳＤⅡ）的重组表达质粒．未经ＩＰＴＧ诱导，该质粒表达的融合蛋白在ＤＨ５α中即有较高的本底表达量．若将带有ＬａｃⅠ基因的ｐＲＥＰ４质粒与重组表达质粒共转化ＤＨ５α菌，... 详细信息

用ｐＧＥＸ载体系统体外构建了一种人精子膜蛋白片段（ＨＳＤⅡ）的重组表达质粒．未经ＩＰＴＧ诱导，该质粒表达的融合蛋白在ＤＨ５α中即有较高的本底表达量．若将带有ＬａｃⅠ基因的ｐＲＥＰ４质粒与重组表达质粒共转化ＤＨ５α菌，则可有效抑制融合蛋白的本底表达．

关键词： pGEX 载体系统精子膜蛋白重组表达质粒

hNOP17,a Novel Binding Protein of hSNF5

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hNOP17,a Novel Binding Protein of hSNF5

The Role of UTX and JMJD3 in Regulating Mashl in RA Induced Neuronal Differentiation of P19 Cells

2008年全国生物化学与分子生物学学术大会

作者： Niu Zhai Ye Zhang Yu-fei Shen (national laboratory of medical molecular biology,department of biochemistry & molecular biology.Institute of Basic medical sciences,chinese academy of medical sciences and peking union medical college,beijing 100005,china)

SWI/SNF complex is one of several evolutionarily conserved multi-protein complexes that remodel chro- matin in an ATP-dependent ***1/SNF5 is one highly conserved subunits of SWI/SNF complex,and it has been found to be...

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The Role of UTX and JMJD3 in Regulating Mashl in RA Induced ...

Regulational Mechanism of SirT Family in Differenciation of Embryonal Carcinoma Cells

2008年全国生物化学与分子生物学学术大会

作者： Jin-po Dai Ye Zhang Yu-fei Shen (national laboratory of medical molecular biology,department of biochemistry and molecular biology,Institute of Basic medical sciences,chinese academy of medical sciences and peking union medical college,beijing,100005,china)

Neuronal differentiation is a complicated process involving strictly regulated temporal and spatial expres- sion of diverse *** regulation including chromatin remodeling and histone modification plays an essential rol...

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Regulational Mechanism of SirT Family in Differenciation of ...

Telomere and telomerase in the initial stage of immortalization of esophageal epithelial cell

2008年全国生物化学与分子生物学学术大会

作者： Jian-yi Lv Ye Zhang Yu-fei Shen (national laboratory of medical molecular biology,department of biochemistry and molecular biology,Institute of Basic medical sciences,chinese academy of medical sciences and peking union medical college,beijing,100005,china)

SirT family,also known as nicotinamide adenine dinuleotide(NAD-)-dependent histion deacetylase(HDAC) are implicated in a variety of cellular functions ranging from gene silencing,over the control of the cell cycle and apoptosis,to energy *** differenciation is a complicated process under diverse regulational ***,the roles of SirT family in the network are sill not fully understand.

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World Journal of Gastroenterology 2002年第2期8卷 357-362页

作者： Zhong-Ying Shen Li-Yan Xu Wei-Jia Cai Min-Hua Chen Jian Shen,department of Tumor Pathology,medical college of Shantou University,Shantou 515031,Guangdong Province,china En-Min Li,department of biochemistry and molecular biology,medical college of Shantou University,Shantou 515031,Guangdong Province,china Yi Zeng,Institute of Virology,chinese academy of Preventive Medicine,beijing 100052,china department of Tumor Pathology Medical College of Shantou University 22 Xinling Road Shantou 515031 Guandong Province China. zhongyingshen@***

AIM: To search for the biomarker of cellular immortalization, the telomere length, telomerase activity and its subunits in cultured epithelial cells of human fetal esophagus in the process of immortalization. METHODS: The transgenic cell line of human fetal esophageal epithelium (SHEE) was established with E(6)E(7) genes of human papillomavirus (HPV) type 18 in our laboratory. Morphological phenotype of cultured SHEE cells from the 6th to 30th passages, was examined by phase contrast microscopy, the telomere length was assayed by Southern blot method, and the activity of telomerase was analyzed by telomeric repeat amplification protocol (TRAP). Expressions of subunits of telomerase, hTR and hTERT, were assessed by RT-PCR. DNA content in cell cycle was detected by flow cytometry. The cell apoptosis was examined by electron microscopy (EM) and TUNEL label. RESULTS: SHEE cells from the 6th to 10th passages showed cellular proliferation with a good differentiation. From the 12th to the 16th passages, many senescent and apoptotic cells appeared, and the telomere length sharply shortened from 23kb to 17kb without expression of hTERT and telomerase activity. At the 20th passage, SHEE cells overcame the senescence and apoptosis and restored their proliferative activity with expression of telomerase and hTERT at low levels, but the telomere length shortened continuously to the lowest of 3kb. After the 30th passage cells proliferation was restored by increment of cells at S and G2M phase in the cell cycle and telomerase activity expressed at high levels and with maintenance of telomere length. CONCLUSION: At the early stage of SHEE cells, telomeres are shortened without expression of telomerase and hTERT causing cellular senescence and cell death. From the 20th to the 30th passages, the activation of telomerase and maintenance of telomere length show a progressive process for immortalization of esophageal epithelial cells. The expression of telomerase may constitute a biomark

关键词： Cell Transformation, Neoplastic Apoptosis Cell Division Cell Line Cell Size Epithelial Cells Esophagus Humans In Situ Nick-End Labeling Papillomavirus, Human Research Support, Non-U.S. Gov't Telomerase Telomere

Establishment of a Screening System for Selection of siRNA Target Sites Directed against Hepatitis B Virus Surface Gene

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Acta Biochimica et Biophysica Sinica 2005年第5期37卷 310-316页

作者： Xiu-Min ZHOU~(1.3) Ju-Sheng LIN~1 Yi SHI~3 De-An TIAN~2 Huan-Jun huang~2 He-Jun ZHOU~1 You-Xin JIN~3 1 Institute of Liver Diseases. Tongji Hospital, Tongji medical college, Huazhong University of Science and Technology, Wuhan 430030, china: 2 department of Gastroenterology, Tongji Hospital, Tongji medical college, Huazhong University of Science and Technology, Wuhan 430030, china 3 State Key laboratory of molecular biology, Institute of biochemistry and Cell biology,Shanghai Institutes for Biological sciences,chinese academy of sciences, Shanghai 200031, china Institute of Liver Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430030 China State Key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China department of Gastroenterology Tongji Hospital Tongji Medical College Huazhong University of Science and TechnologyWuhan 430030 China

To study the inhibitory effects of plasmid-derived small interfering RNA(siRNA)andsynthetic siRNA on the expression of the hepatitis B virus surface(HBs)gene,three plasmid-derived siRNAsand one synthetic siRNA that complement the coding region of the HBs gene were *** HBsexpression plasmid pHBs-EGFP was also *** cells were co-transfected with pHBs-EGFPand the above *** HBs mRNA quantities were measured by reverse-transcription PCR,and thelevel of HBs-EGFP fusion protein was quantified by fluorescent *** concentrations of thehepatitis B virus surface antigen(HBsAg)derived from the culture supernatant of transfected HepG2.2.15cells were measured by an enzyme-linked immunosorbent assay(ELISA)*** results showed that thethree plasmid-derived siRNAs and the synthetic siRNA can effectively reduce the quantities of HBs mRNAand *** plasmid-derived siRNA psiRNA1 was found to be the most effective inhibitor of *** can inhibit HBs-EGFP expression by 63.3% and suppress HBs mRNA by 75.6%.To furthersubstantiate the above observations,psiRNA1 was transfected into HepG2.2.15 cells(an HBV secreting cellline).The transfections resulted in almost complete blockage of HBsAg production,whereas control vector-transfected cells secreted high levels of HBsAg 7 days *** conclusion,our data suggeststhat RNA interference(RNAi)is an efficient approach for reducing the level of HBs transcripts and proteinsand for suppressing HBsAg production.

关键词： RNA interference(RNAi) small interference RNA(siRNA) hepatitis B virus surface gene(HBs gene)

编码内质网膜蛋白的新基因家族RTN的研究进展

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生物化学与生物物理进展 2001年第1期28卷 40-43页

作者：黄晓玮阴彬袁建刚强伯勤中国医学科学院基础医学研究所中国协和医科大学基础医学院医学分子生物国家重点实验室北京100005 国家人类基因组北方研究中心北京100073

蛋白质的分泌渠道是近几年生命科学研究焦点之一 .RTN (reticulon)是定位于内质网膜上的一个基因家族 ,现已发现RTN1、RTN2、RTN3、RTN4等多个家族成员 .其中 ,这些基因在蛋白质分泌加工过程中的具体作用 ,以及RTN1(即NSP)作为神经内分... 详细信息

蛋白质的分泌渠道是近几年生命科学研究焦点之一 .RTN (reticulon)是定位于内质网膜上的一个基因家族 ,现已发现RTN1、RTN2、RTN3、RTN4等多个家族成员 .其中 ,这些基因在蛋白质分泌加工过程中的具体作用 ,以及RTN1(即NSP)作为神经内分泌特异性的标志或与神经元分化的关系都在进一步探讨之中 .

关键词：内质肉膜蛋白蛋白质分泌神经内分泌特异蛋白分泌渠道