狂犬病病毒核蛋白基因在大肠杆菌中的高效表达

为获得狂犬病病毒(RV)核蛋白抗原,采用RT-PCR方法从狂犬病病毒ERA株中扩增了RV核蛋白基因,分别亚克隆到原核表达载体pET-28a(+)和pET-32a(+)中,经PCR和双酶切鉴定以及序列测定,重组质粒构建成功。将重组质粒转化到大肠杆菌BL21(DE3)中进行表达,表达的核蛋白再经Ni2+-NTA亲和层析纯化。结果显示,核蛋白在pET-32a(+)中的表达量(30.4%)高于在pET-28a(+)中的表达量(19.4%),培养物中的高纯度核蛋白产量分别为13.6和8.45 mg/L。经Western-blotting检测,不同载体表达的核蛋白均可被兔抗RV多克隆抗体特异识别,表明核蛋白具有良好的反应原性。     

Nonparticipation as Self-Censorship: Publicly Observable Political Activity in a Polarized Opinion Climate

In a polarized opinion climate, people may refrain from participating in publicly observable political activities that make them vulnerable to scrutiny and criticism by others who hold opinions that differ from their own. We took a dispositional approach to testing this claim by determining whether people who are relatively more influenced by the climate of opinion when choosing whether or not to voice an opinion, measured with the Willingness to Self-Censor scale [Hayes et al. International Journal of Public Opinion Research 17 (2005) 298], are also relatively less likely to engage in public political activities. In a poll of residents of the United States, we found that even after controlling for interest in politics, political ideology, ideological extremity, political efficacy, attention to political news, dispositional shyness, frequency of political discussion, and demographics, dispositional self-censors reported having engaged in relatively fewer public political activities over the prior 2 years compared to those less willing to censor their own opinion expression. These results are consistent with our interpretation of political participation as a social process that is governed in part by the social psychological implications of participation to the person. At a larger theoretical level, our findings connect the literature on opinion perceptions and opinion expression with research on political participation.

社会转型期公众民意表达科学化的理性思考

当前,我国正处于社会转型深化期和攻坚期,公众民意表达发生了深刻变化,呈现出利益诉求不断扩大的趋势,突出表现为多元化、差异化和群体化等新特点。在面对公众民意表达时,政府与公众应在公共理性的框架内,寻求理性和有序的解决途径。为提升公众民意表达的科学化水平,必须加强和创新社会管理,大力营造党委、政府、社会和公众理性互动的社会管理环境,创新发展治理型的社会管理制度,不断拓宽政府主导的规范高效、理性有序的社会管理路径。     

浅谈电视节目模式的著作权保护

随着互联网的发展和观众需求的日益多样化,全球范围内的电视节目模式许可交易日益增长,同时也产生了诸多法律纠纷,而电视节目模式的法律保护仍然存在立法空白。结合欧美法院判例和学术界的观点,本文对中美电视节目模式的法律保护现状、电视节目模式的侵权认定以及在现阶段如何对电视节目模式进行法律保护作出了初步探讨,得出结论是未来趋势应将电视节目模式作为汇编作品予以著作权法的保护,这也是建设社会主义文化强国,增强国家文化软实力的必要举措。     

形象权转换性使用规则研究

未经许可借用他人形象制作虚拟人物角色和动漫表情包的现象越来越普遍,应该完全保护个人的肖像权还是为公众自由创作虚拟人物形象和表情包留出空间值得思考。形象权制度下的一系列判例借用版权合理使用制度中的转换性使用规则平衡形象权保护和公众的表达自由,能为如何平衡个人的肖像保护和公众的参与创作和表达带来借鉴。本文通过分析转换性使用规则在形象权案例中的运用,提出应调整转换性使用规则以增强适用该规则判定形象权案例的稳定性。     

NF-κB受体激活因子配体的表达及其多克隆抗体的制备

为获取猪源NF-k B受体激活因子配体(RANKL)及其抗体,根据GenBank中猪源RANKL的序列设计引物,将合成的RANKL基因克隆至pMAL-C4x表达载体,转化到大肠杆菌DH5α。经PCR和双酶切鉴定后测序分析,将阳性克隆转化至大肠杆菌BL21后通过IPTG诱导表达。优化表达条件,确定是包涵体表达,蛋白大小为78.1 ku。使用纯化的RANKL为抗原免疫兔制备多克隆抗体。构建重组质粒pCMV-RANKL,表达目的蛋白并使其与多克隆血清反应。经ELISA、Western-blot和IFA鉴定,证实抗RANKL多克隆抗体有良好的抗原特异性,为后续黏膜免疫的研究奠定了基础。     

西尼罗病毒E蛋白的原核表达及其多克隆抗体的制备

利用PCR技术扩增西尼罗病毒(WNV)E蛋白基因并将其插入原核表达载体pET-30a(+),构建重组质粒pET-30a(+)-WNV-E,并转化至大肠杆菌BL21(DE3)感受态细胞中,经IPTG诱导目的蛋白表达,对诱导条件进行优化,利用His镍柱亲和层析法纯化目的蛋白,纯化后的蛋白经SDS-PAGE与Westernblot鉴定正确后,免疫BALB/c小鼠制备多克隆抗体并进行间接免疫荧光鉴定。结果显示:WNV E蛋白在大肠杆菌中以包涵体形式被成功表达,蛋白最佳诱导条件为37℃,0.8 mmol/L IPTG诱导5 h,经间接免疫荧光鉴定多克隆抗体能特异性识别WNV E蛋白。本研究成功表达并纯化了WNV E蛋白,证明该蛋白具有良好的免疫原性,为WNV抗原、抗体检测方法的建立和新型疫苗的研发及相关研究奠定了基础。     

大鼠急性肾缺血再灌注损伤早期的基因表达

目的研究急性肾缺血再灌注损伤早期的差异基因表达,探索其潜在的分子机制。方法通过单纯夹闭大鼠肾动脉制备肾缺血再灌注模型,进行基因表达芯片检测和生物信息学分析,筛选急性肾损伤早期的差异基因表达及其所涉及的细胞活动和信号通路。同时,通过构建差异表达基因的生物网络来寻找与急性肾损伤关系密切的分子,并进行荧光定量PCR验证。结果在检测出的151个差异表达基因中,132个基因显著上调,19个基因显著下调,GO与KEGG通路富集分析结果显示主要与细胞增殖、细胞因子介导的信号通路和免疫反应等有关。荧光定量PCR结果显示,与对照组相比,挑选出的3个与急性肾损伤高度相关的基因(ANXA1、PHLDA1、KLF6)在疾病早期具有显著的表达差异,上升倍数与芯片检测出的倍数基本一致。结论本研究揭示了急性肾损伤早期的差异基因表达特征以及所涉及的生物学过程和信号通路,其中ANXA1、PHLDA1、KLF6可能在急性肾损伤的发病机制中起到一定作用。     

A Tale of Two Leaders: Leadership and Cultural Change at the New Zealand Treasury

Differences in the transformational leadership exercised at the New Zealand Treasury (NZT) by two Secretaries – Henry Lang (1969-1973) and Graham Scott (1986-1996) – are related to differences in their paradigmatic situation rather than a shift away from a serial loyalist public service bargain that caused both to direct their leadership initiatives toward developing and maintaining the trust successive finance ministers placed in NZT advisors. Thus while Lang's leadership was directed toward expanding the reporting role of the NZT to the limits of what could be expected under a relatively stable market failure policy paradigm, the subsequent erosion of the authority of this paradigm meant that, under Scott's leadership, the NZT reinvented itself to avoid implication in a failure to contribute to the policy leadership required to advance a coherent reform strategy.     

韩国国家安全与表达自由案例研究

国家安全与表达自由之间的冲突是每一个国家在任何一个历史阶段均必须直面的重大问题。韩国专制主义盛行的20世纪60年代至90年代中期所涉及的“国家安全与表达自由”典型司法案例反映了韩国普通法院及宪法法院在衡平这两者价值间冲突的立场变迁及韩国法院在维护人权和促进韩国民主与法制发展中所扮演的角色,可为中国学界在“国家安全与表达自由”这样复杂而又棘手的问题上,提供些许可借鉴的外域经验。