共查询到20条相似文献，搜索用时 15 毫秒
1.
Li D Wang QQ Tang GP Huang HL Shen FP Li JZ Yu H 《Journal of Zhejiang University. Science. B》2006,7(11):906-911
Objective： To construct a novel kind of nonviral gene delivery vector based on polyethylenimine （PEI） conjugated with polypeptides derived from ligand FGF with high transfection efficiency and according to tumor targeting ability. Methods： The synthetic polypeptides CR16 for binding FGF receptors was conjugated to PEI and the characters of the polypeptides including DNA condensing and particle size were determined. Enhanced efficiency and the targeting specificity of the synthesized vector were investigated in vitro and in vivo. Results： The polypeptides were successfully coupled to PEI. The new vectors PEI-CR16 could efficiently condense pDNA into particles with around 200 nm diameter. The PEI-CR16/pDNA polyplexes showed significantly greater transgene activity than PEI/pDNA in FGF receptors positive tumor cells in vitro and in vivo gene transfer, while no difference was observed in FGF receptors negative tumor cells. The enhanced transfection efficiency of PEI-CR 16 could be blocked by excess free polypeptides. Conclusion： The synthesized vector could improve the efficiency of gene transfer and targeting specificity in FGF receptors positive cells. The vector had good prospect for use in cancer gene therapy. 相似文献
2.
《天津大学学报(英文版)》2015,(3)
In order to modify inorganic particles as chromatic electrophoretic particles, an approach was designed and used to prepare Fe2O3 red electrophoretic particles. These Fe2O3-cationic hybrid nanoparticles(Fe2O3-CHNPs)were prepared through Fe2O3 core covered with polymer shell which was composed of SiO 2 and P(DMAEMA-co-HMA)by using atom transfer radical polymerization(ATRP)technique. The Si O2-coating could introduce the functional group on the surfaceof inorganic particles, through which the polymer shell could be formed by using ATRP technique. The results of Fourier transform infrared spectra(FT-IR), X-ray photoelectron spectroscopy(XPS)and thermal gravimetric analysis(TGA)confirmed the chemical compositions of Fe2O3-CHNPs; the images of transmission electron microscopy(TEM)indicated the core-shell structure of Fe2O3-CHNPs; the measurements of dynamic light scattering(DLS)showed a 253.7 nm average particle size with narrow size distribution; and the zeta potential measurements identified the high chargeability of Fe2O3-CHNPs. Furthermore, the resulting nanoparticles were successfully applied in the electrophoretic display cell, which demonstrated that it was an effective approach to preparing chromatic electrophoretic particles. 相似文献
3.
Objective: To construct a eukaryotic expression plasmid pcDNA3.1 (-)-Humanin. Methods: The recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1 (-) and the recombinant plasmids pcDNA3, l(-)-Humanin were identified by sequencing. Results: Recombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence. Conclusions: A eukaryotic expression plasmid of Humanin was successfully constructed. 相似文献
4.
饱和苯酚氯仿法提取Tg2576转基因鼠基因组DNA,PCR法扩增编码β-淀粉样蛋白的目的基因,利用基因克隆技术构建以β-淀粉样蛋白为靶的表达载体pcDNA3.1-Aβ42×2,应用酶切及测序鉴定表达载体.相应的双酶酶切能够获得插入的目的基因片段(为269bp),测序未发现突变,表达载体构建成功. 相似文献
5.
增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),也称周期蛋白或DNA聚合酶的辅助蛋白,是真核细胞合成所必需的核蛋白,在DNA复制中起重要作用。前期实验发现日本七鳃鳗肝脏cDNA文库的表达序列标签(Expressed Sequence Tag,EST)中存在与高等脊椎动物pcna基因同源的序列。提取日本七鳃鳗(Lampetra japonica)肝脏组织RNA,通过RT-PCR方法扩增七鳃鳗pcna基因,对其进行生物信息学分析,并将Lj-pcna基因成功构建到pGFP-N2真核表达载体上,重组质粒PGFP-N2-Lj-pcna转染人Hela细胞,荧光显微镜下观察有荧光蛋白的表达。日本七鳃鳗pcna基因的真核表达载体成功构建和转染,为探讨七鳃鳗pcna基因功能研究及其它七鳃鳗相关研究提供条件。 相似文献
6.
刘娟 《湖北广播电视大学学报》2011,31(11):159-160
实验测试了氨基磺酸系高效减水剂AH溶液的表面张力、AH在水泥颗粒表面的吸附量及ξ电位,结果表明：AH能够减小溶液表面张力,使分散体系的自由能降低;它在水泥颗粒表面呈环圈及尾状吸附,产生较大的立体空间位阻;掺AH的水泥粒子表面ξ电位绝对值较大,水泥颗粒间存在着较强静电斥力。这些均使得AH具有良好分散作用。此外,AH分子结构中存在的极性基团与水分子间的氢键缔合,在水泥粒子表面形成的溶剂化水膜的润滑,也对AH分散起到一定的加强作用。 相似文献
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8.
1IntroductionRibosome-inactivating proteins(RIPs)occur natu-rally in a variety of higher plant species,and theyfunction by catalytic depurination of a specific aden-osine residue located near the3*terminus of eukary-otic large ribosomal subunit rRNA,preventing EF-2/GTP binding and thereby blocking peptidyl-tRNAtranslocation during protein synthesis[1].Many RIPsare potent antiviral and antifungal proteins in vitro[2,3],but it may beinsufficient for field application,as com-pared to the ac… 相似文献
9.
The reaction of ZnO nanoparticles grafted with KH570 silane coupling agent was carried out in water-alcohol mixed solvent. Several characterization methods were applied to analyzing the results of surface modification such as Soxhlet extraction, Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), thermogravimetry (TG), differential scanning calorimetry (DSC) and zeta potential (ζ). The results of FT-IR and TG-DSC show that the desired reaction chains have been covalently bonded on the surface of ZnO nanoparticles. Zetasizer results reveal that themaximal absolute value of ζ of the modified ZnO particles in acetone medium was 67.0 mV, which was much higher than that of the unmodified ZnO particles. So the surface of nanosized ZnO changed from hydrophilicity to hydrophobicity and the dispersity of ZnO nanoparticles were improved simultaneously. Finally, the mechanism of graft modification was discussed. 相似文献
10.
利用化学共沉淀法制备了磁性Fe3O4纳米微粒,用硅烷偶联剂MPS对所制备的磁性微粒进行表面有机改性,并用FTIR、XRD、TEM、XPS等表征方法对样品进行了表征,结果表明:MPS已经很好地键连到了磁性Fe3O4纳米微粒的表面。改性的磁性Fe3O4纳米微粒具有亲水和亲油两种性质,采用改性后的磁性微粒可以显著改善磁性微球的性能指标。 相似文献
11.
从假单胞菌（Pseudomonassp．）XZG36中克隆弹性蛋白酶基因,构建原核表达载体,实现其在大肠杆菌（Escherichiacoli）中的高效表达,并对表达产物进行酶学性质分析,为微生物发酵生产弹性蛋白酶奠定基础．以假单胞菌基因组DNA为模板,PCR扩增弹性蛋白酶基因,并将其开放阅读框（0RF）克隆至融合表达载体pET30a（＋）进一步IPTG诱导表达;表达产物经His·Bind亲和层析纯化后对弹性蛋白酶进行酶学性质分析．实验成功克隆了弹性蛋白酶基因,DNA基因片段为1672bp、编码497个氨基酸残基的多肽,与预计长度相符合;实现了其在E．coli中的高效表达,表达量约占菌体总蛋白的20％;经SDS-PAGE分析,相对分子质量为48000,与预期的一致;提纯后的表达蛋白SDS-PAGE分析可见单一条带,纯度可达92％以上．表达蛋白具有良好活性． 相似文献
12.
Fu Y Wang SQ Liu YP Wang GP Wang JT Gong SS 《Journal of Zhejiang University. Science. B》2008,9(4):299-305
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 successfully 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. 相似文献
13.
In this study, the barite nanoparticles were successfully modified with stearate and the influence of stearate addition on
the performance of barite nanoparticles was systematically investigated. The products were characterized by activating factor
analysis, contact angle test, surface energy calculation, sedimentation rate calculation, rheological measurement, and FT-IR
analysis, etc. As the quantity of added stearate increased, both the activating factor and contact angle of barite nanoparticles increased
first then decreased. When the stearate content was 5% of the mass of barite nanoparticles, the activating factor and water
contact angle of modified particles reached maximum value, 97% and 126° respectively. At this time, the sedimentation rate
reached minimum, and so did the surface energy. The rheological test reveals that the viscosity of modified barite nanoparticles/petronol
system decreases greatly, indicating the surface performance of barite nanoparticles has changed from hydrophilicity to lipophilicity
after modification. C=O and COO stretching vibration peaks were found in the FT-IR spectra, which proves that the stearate
has combined onto the surface of barite nanoparticles. Finally, according to the zeta potential result of unmodified barite,
the possible modification mechanism was provided.
Project supported by the International Corporation Project (Grant No.07SU07001), and the Science Foundation of Science and
Technology Commission of Shanghai Municipality (Grant No.0552nm011) 相似文献
14.
利用化学共沉淀法制备了磁性Fe3O4纳米微粒,用硅烷偶联剂MPS对所制备的磁性微粒进行表面有机改性,并用FTIR、XRD、TEM、XPS等表征方法对样品进行了表征,结果表明:MPS已经很好地键连到了磁性Fe3O4纳米微粒的表面。改性的磁性Fe3O4纳米微粒具有亲水和亲油两种性质,采用改性后的磁性微粒可以显著改善磁性微球的性能指标。 相似文献
15.
Clinical features of chronic hepatitis B patients with YMDD mutation after lamivudine therapy 总被引：5，自引：0，他引：5
INTRODUCTION Chronic hepatitis B(CHB)remains a major public health problem,affecting more than350mil-lion people worldwide.Cirrhosis,liver failure,or hepatocellular carcinoma will develop in approxi-mately15to40percent of infected patients(Ganem and Prince,2004).Lamivudine,an oral nucleoside analogue,inhibits HBV replication(de Clercq,2001;2004;Lai et al.,1997;1998;Marcellin et al.,2004).It can markedly reduce serum HBV DNA levels and normalize alanine aminotransferase(ALT)levels … 相似文献
16.
Polyethylenimine-cyclodextrin-tegafur (PEI-CyD-tegafur) conjugate was synthesized as a novel multifunctional prodrug of tegafur
for co-delivery of chemotherapeutic agent tegafur and enhanced green fluorescent protein (EGFP) reporter plasmid DNA. Conjugation
of tegafur to PEI-CyD via chemical linkage was characterized by 1H NMR spectrometry and ultraviolet (UV) spectrometry. PEI-CyD-tegafur was able to condense plasmid DNA into complexes of around
150 nm with positive charge at the N/P ratio of 25, in accordance with electron microscopy observation of compact and monodisperse
nanoparticles. The results of in vitro experiments showed enhanced cytotoxicity and considerable transfection efficiency in
B16F10 cell line. Therefore, PEI-CyD-tegafur may have great potential as a co-delivery system with anti-cancer activity and
potential for gene delivery. 相似文献
17.
目的:在大肠杆菌(BL21)中构建可溶性表达的金黄色葡萄球菌B型肠毒素(SEB)受体拮抗剂。方法:首先确定SEB受体桔抗剂的基因序列,然后用含有SEB受体桔抗剂的基因序列重组质粒表达载体PGEX-4T-1转化大肠杆菌BL21(DE3),利用IPTG诱导表达获得蛋白,产物经GST柱纯化后,利用ELISA检测其与SEB的结合能力并进行其体内外药效学实验。结果:该质粒成功转化为可溶性表达,ELISA结果显示表达产物可与SEB特异性结合。结论:本研究成功对SEB受体拮抗剂GST可溶性表达并对其活性进行初步分析。 相似文献
18.
根据已发表的序列，通过PCR技术克隆了一系列构建烟草叶绿体多顺反子表达载体所需的元件：质体核搪体（16S）RNA操纵元启动子（Prrn）、质体面A基因3’端（psbA3’）、山菠菜甜菜碱醛脱氢酶基因（BADH）、烟草叶绿体高频同源重组片段（psaA／psbC，大小3463bp）、甘露聚糖酶基因（man）、绿荧光蛋白基因（gfp）。构建了烟草质体多顺反子定点整合表达载体pLM7（-psaA-Prrn-SD-man-SD-gfp-SD-BADH-PSBA3’-PSBC-）。并在大肠杆菌中通过平板定性分析等方法对所构建载体上的表达盒进行了功能鉴定。 相似文献
19.
乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)的慢性感染,与慢性病毒性肝炎、肝硬化、肝细胞癌(HCC)的发病密切相关,其发病机理涉及到很多基因的共同参与.病毒基因的复制和表达受到肝细胞中蛋白质因子的调节,肝炎病毒基因编码的蛋白与肝细胞中的蛋白能够结合,肝炎病毒蛋白在肝细胞中的表达对于肝细胞的基因表达谱产生影响,也可能是病毒性肝炎、肝硬化、肝细胞癌发病机制的重要机制.酵母单杂交技术、酵母双杂交技术、基因芯片技术、抑制性消减杂交技术、噬菌体展示技术、蛋白质分离纯化与基因克隆化的反向遗传学技术等,在肝炎病毒基因调控、肝炎病毒蛋白结合蛋白的研究、肝炎病毒蛋白反式激活靶基因的研究中都有重要的应用,是促进慢性病毒性肝炎发病机理研究,探索病毒性肝炎新型治疗技术和治疗方法的有效途径. 相似文献