检索结果-南通市图书馆

Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein

引用

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2008年第4期9卷 299-305页

作者： Yong FU Shen-qing WANG Ying-peng LIU Guo-peng WANG Jian-ting WANG Shu-sheng GONG Department of Otorhinolaryngology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou 310003 China Department of Otorhinolaryngology Union Hospital of Tongii Medical College Huazhong University of Science and Technology Wuhan 430022 China Department of Surgery-Otolaryngology University of Kentucky Medical Center Lexington Kentucky 40536 USA Department of Surgery-Otolaryngology Beijing Tongren Hospital Capital Medical University Beijing 100730 China

Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. Methods: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. Results: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. Conclusion: We have success- fully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies.

关键词： Recombinant adenovirus vector Viral infection Fetal neural stem cells Green fluorescent protein

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Transcriptional Regulation of Urokinase Receptor in High-(95D) and Low-metastatic (95C) Human Lung Cancer Cells

引用

Acta Biochimica et Biophysica Sinica 2004年第6期36卷 405-411页

作者： Chun-Yang LI, Li TAN, Guo-Jing ZHANG, Ping LI, Chang TONG, Jing FAN, and Yun-Song ZHU* Key Laboratory, the Education Ministry, Department of Biochemistry and Molecular Biology, Shanghai Medical College,Fudan University, Shanghai 200032, china Key Laboratory the Education Ministry Department of Biochemistry and Molecular Biology Shanghai Medical College Fudan UniversityShanghai 200032 China

To study the transcriptional regulation of urokinase receptor (uPAR) in high- (95D) and low-metastatic (95C) human lung cancer cells, we performed PCR to amplify 2238 bp uPAR promoter from 95C and 95D cells. According to the results of sequencing, five different bases are found in uPAR promoter between 95C and 95D cells. The results of luciferase activity assay showed that these differences have no significant effect on the uPAR promoter activity. Based on a normal uPAR promoter, progressive truncated mutants were constructed. The transient transfection/reporter assay showed that the promoter region from –136 to +9 may interact with relevant nuclear factors, which result in different levels of uPAR expression between 95C and 95D cells.

关键词： uPAR promoter transcriptional regulation

来源：

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Influence of Trace Amount of Calponin on Smooth Muscle Myosin in Different States

引用

Acta Biochimica et Biophysica Sinica 2004年第8期36卷 529-536页

作者： Jing-Xian YANG, Xiao-Hua FENG, Ying ZHANG, and Yuan LIN Department of Pharmacology, Dalian Medical University, Dalian 116027, china Department of Pharmacology Dalian Medical UniversityDalian 116027China

Calponin (CaP), a thin filament-associated protein, is thought to be involved in modulating smooth muscle contractile activity, but the role and mechanism keep unknown. In this study, trace amount of calponin (TAC) was found to obviously influence myosin in different states in Ca2+-independent manner, suggesting a high efficient interaction between TAC and myosin. In this assay, the lowest ratio of CaP vs. myosin was 1:10,000, with the concentration of CaP 10,000-fold lower than that used previously. Myosin phosphorylation, myosin Mg2+-ATPase activity and protein binding activity were detected to determine the effects of TAC on the myosin in different states. The amount of precipitated myosin that bound to TAC was used as the index to determine the interaction between myosin and TAC in binding assay. Our data showed that in the absence of actin, TAC significantly increased the precipitation of unphosphorylated myosin, Ca2+- dependently or independently phosphorylated myosin by MLCK, and stimulated the Mg2+-ATPase activities of these myosins slightly but significantly. However, no obvious change of precipitation of myosin phospho- rylated by PKA was observed, indicating the relatively selective effect of TAC. In the presence of actin, the increase of myosin precipitations was abolished, and no obvious change of actin precipitations and actin- activated myosin Mg2+-ATPase activities were observed implicating the high efficiency of TAC on myosin being present in the absence of actin. Although we can not give conclusive comments to our results, we propose that the high efficiency of TAC-myosin interaction is present when actin is dissociated from myosin, even if CaP/myosin ratio is very low; this high efficient interaction can be abolished by actin. However, why and how TAC can possess such a high efficiency to influence myosin and how the physiological significance of the high efficiency of TAC is in regulating the interaction between myosin and actin remain to be inve

关键词： trace amount of calponin protein binding phosphorylation myosin Mg2+-ATPase

来源：

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

The limit analysis in soil and rock:a mature discipline of geomechanics

引用

Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2007年第11期8卷 1712-1724页

作者： CHEN Zu-yu china Institute of Water Resources and Hydropower Research Beijing 100044 China MOE Key Laboratory of Soft Soils and Geoenvironmental Engineering Zhejiang University Hangzhou 310027 China

The solution of a slope stability problem can be approached by its least upper-bound and maximum lower-bound with high accuracy. The limit equilibrium methods that employ vertical slices imply a lower bound of the factor of safety. It has been successfully extended to the area of active earth pressure analysis that accounts for different input of locations of earth pressure applications. Those methods that employ slices with inclined interfaces give an upper-bound approach to the stability analysis. It enjoys a sound mechanical background and is able to provide accurate solutions of soil plasticity. It has been successfully extended to the area of bearing capacity analysis in which various empirical coefficients are no longer necessary. The 3D upper- and lower-bound methods under this framework have been made possible and show great potential for solving various engineering problems.

关键词： Limit analysis Geomechanics Slope stability problem Limit equilibrium method

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Isolation and Partial Characterization of a Novel Pollen-specific cDNA with Multiple Polyadenylation Sites from Wheat

引用

Acta Biochimica et Biophysica Sinica 2004年第7期36卷 467-476页

作者： Yong-Feng JIN* and Teng-Fei BIANInstitute of Biochemistry, Zhejiang University, Hangzhou 310029, china Institute of Biochemistry Zhejiang University Hangzhou 310029 Chin

A novel pollen-specific full-length cDNA clone PSG076 was isolated using suppression subtractive hybridization and 5’/3’ RACE techniques. PSG076 was shown to exhibit multi-site polyadenylation by sequencing the 3’ ends of the cDNAs. At least six transcripts with different length were produced from the single gene based on different poly(A) tail attachment sites. However, polyadenylation consensus sequence AAUAAA was not seen at the 3’-untranslated sequence. PSG076 contained a 299 bp 5’ untranslated region and an open reading frame of 663 bp encoding a 221 amino acid peptide with pI of 4.31. A blast search revealed that this sequence did not show a significant similarity to any genes deposited in the public database. Southern blot indicated that PSG076 was a single copy gene. Northern blot and RT-PCR analysis indicated that PSG076 transcripts showed specific expression in mature pollen, and weak or undetectable signals in uninucleate microspore, immature seed, stem, young leave, root and ovary. Further analysis of the expression pattern in gametophyte showed that PSG076 transcripts were undetectable in uninucleate, binucleate microspore and pollen at early stage, and were first detectable and increased rapidly at middle and late stages of pollen development with the maximum level in mature pollen and also expressed in germinating pollen in vivo, suggesting that PSG076might play a role in pollen germination and pollen tube growthin addition to its function in maturation. The evidences gathered in this work indicated that the six different transcripts from the single gene were differentially expressed during pollen development.

关键词： wheat (Triticum aestivum L.) pollen-specific cDNA suppression subtractive hybridization multiple-site polyadenylation differential expression

来源：

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Magnetic field and performance analysis of a tubular permanent magnet linear synchronous motor applied in elevator door system

引用

Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2008年第4期9卷 572-576页

作者： Xiao LIU Yun-yue YE Zhuo ZHENG Qin-fen LU School of Electrical Engineering Zhejiang University Hangzhou 310027 China

A novel elevator door driven by tubular permanent magnet linear synchronous motor (TPMLSM) is presented. This TPMLSM applies axial magnet array topology of the secondary rod, air-cored armature windings and slotless structure of the forcer to improve the stability of the thrust. The influence of two major dimensions, the pitch and radius of the permanent magnet (PM), on magnetic field was studied and the best values were given by the finite element analysis (FEA). The magnetic field, back EMF and thrust of the motor were analyzed and the PM size was optimized to reduce the harmonic components of the magnetic field and improve the performance of the motor. Predicted results are validated by the experiment. It is shown that the performance of the motor and the novel elevator door system is satisfying.

关键词： Elevator door system Tubular permanent magnet linear synchronous motor (TPMLSM) Finite element analysis(FEA)

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Cloning, expression, purification, and characterization of LC-1 ScFv with GFP tag

引用

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2005年第8期6卷 832-837页

作者：卢敏龚兴国于红郦剑勇 Institute of Biologic Macromolecule and Enzyme Engineering School of Life Science Zhejiang UniversityHangzhou 310027 China

Total RNA was isolated from the hybridoma cell line （LC- 1 ）, which secretes anti-lung adenocarcinoma monoclonal antibody, and was transferred into cDNA. Based on the FRl （framework region l） and FR4 conserved regions of LC-1 gene, the variable regions of heavy chain （Vh） and light chain （Vl） were amplified, and the Vh and modified Vl were connected to single chain Fv （ScFv） by SOE-PCR （splice overlap extension PCR）. The modified ScFv was fused with green fluorescent protein （GFP） and introduced into E. coli JM109. The fusion protein induced by lPTG （Isopropylthiogalactoside） was about 57000 on a 10% SDS-PAGE gel （10% Sds Polyacrylamide Gel Electrophoresis）, and primarily manifested as inclusion bodies. The renatured protein purified by Ni-NTA Superflow resins showed ability to bind to antigen on SPC-A-l lung adenocarcinoma. In addition, the induced host cells fluoresced bright green under 395 nm wavelength, which indicated that the expected protein with dual activity was expressed in the prokaryotic system. The ScFv with GFP tag used in this research can be applied as a new reagent to detect immunological dye, and provide a feasible way to detect adenocarcinoma in a clinical setting.

关键词： ScFv (single chain variable fragment) GFP (green fluorescent protein) tag Protein fusion Purification

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

A Novel Gene Delivery System Targeting Urokinase Receptor

引用

Acta Biochimica et Biophysica Sinica 2004年第7期36卷 485-491页

作者： Xing-Hui SUN, Li TAN, Chun-Yang LI, Chang TONG, Jin FAN, Ping LI, and Yun-Song ZHU*Department of Molecular Genetics, Key Laboratory of Molecular Medicine of Ministry Education,Shanghai Medical College of Fudan University, Shanghai 200032, china Department of Molecular Genetics Key Laboratory of Molecular Medicine of Ministry Education Shanghai Medical College of Fudan University Shanghai 200032 China

Recombinant proteins that combine different functions required for cell targeting and intra- cellular delivery of DNA present an attractive approach for the development of nonviral gene delivery vectors. Here, we described a novel protein termed ATF-lys10 which facilitated cell-specific gene transfer via receptor-mediated endocytosis. ATF-lys10 was composed of the amino-terminal fragment of urokinase and ten lysines at the carboxyl terminus. Bacterially expressed ATF-lys10 protein existed in soluble form, and had antigenicity of human urokinase. Purified ATF-lys10 specifically bound to uPAR-expressing cells and formed protein-DNA complexes with plasmid pGL3-control. After neutralization of excess negative charge with poly-L-lysine, these complexes served as a specific gene delivery vector for uPAR-expressing cells. Lyso- somotropic compounds, such as chloroquine, drastically increased the ATF-lys10 mediated gene delivery efficiency. Our results suggest that the recombinant protein ATF-lys10 with the properties of DNA binding and tumor cell targeting represents a promising method for gene transfer and expression in tumor cells.

关键词： amino-terminal fragment urokinase gene transfer endocytosis urokinase receptor

来源：

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Removal of hexavalent chromium from aqueous solution by iron nanoparticles

引用

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2005年第10期6卷 1022-1027页

作者：牛少凤刘永徐新华楼章华 Department of Environmental Engineering Zhejiang University Department of Environmental Engineering Zhejiang University Hangzhou 310027 China School of Civil Engineering and Architecture Zhejiang University Hangzhou 310027 China

Groundwater remediation by nanoparticles has received increasing interest in recent years. This report presents a thorough evaluation of hexavalent chromium removal in aqueous solutions using iron (Fe0) nanoparticles. Cr(VI) is a major pollutant of groundwater. Zero-valent iron, an important natural reductant of Cr(VI), is an option in the remediation of contami- nated sites, transforming Cr(VI) to essentially nontoxic Cr(III). At a dose of 0.4 g/L, 100% of Cr(VI) (20 mg/L) was degraded. The Cr(VI) removal efficiency decreased significantly with increasing initial pH. Different Fe0 type was compared in the same con- ditions. The reactivity was in the order starch-stabilized Fe0 nanoparticles>Fe0 nanoparticles>Fe0 powder>Fe0 filings. Electro- chemical analysis of the reaction process led to the conclusion that Cr(OH)3 should be the final product of Cr(VI). Iron nanopar- ticles are good choice for the remediation of heavy metals in groundwater.

关键词： Hexavalent chromium, Iron nanoparticles, Starch, Stabilize, Remediation

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Mechanical design and locomotion control of a homogenous lattice modular self-reconfigurable robot

引用

Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2006年第3期7卷 368-373页

作者：夏平朱新坚费燕琼 School of Electronics and Information Technology Shanghai Jiao Tong University Shanghai 200030 China Research Institute of Robotics Shanghai Jiao Tong University Shanghai 200030 China

In this paper, we propose a novel, 3D, like cubic shape, modular self-reconfigurable (MSR) robot named M-Cubes. Its key mechanical components are analyzed in detail. By communicating with the neighboring modules, each unit employs its automatic lock device composed of a pin and a hole on each connection plane which can connect or disconnect with neighboring modules. The M-Cubes system consisting of many identical modules cooperates to change their connection, and then the whole structure transforms into arbitrary structure. Furthermore, we describe its locomotion control based on the driving function and the adjacency matrix which is effective for solving the computationally difficult problem and optimizing the system motion path during the self-reconfiguration process. Finally, a simulation experiment using java 3D technology, proved the new method for controlling modular robot is robust and useful.

关键词： Robot Module Self-reconfigurable Locomotion control

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：