Peptide aptamers: exchange of the thioredoxin-A scaffold by alternative platform proteins and its influence on target protein binding

Peptide aptamers have emerged as powerful new tools for molecular medicine. They can specifically bind to and functionally inactivate a given target molecule under intracellular conditions. Typically, peptide aptamers are generated by screening a randomized peptide expression library, displayed from the Escherichia coli thioredoxin A (TrxA) protein. Here, we transferred peptide moieties from defined TrxA-based peptide aptamers to alternative scaffold proteins, such as the green fluorescent protein and staphylococcal nuclease. Yeast and mammalian two-hybrid assays as well as in vitro binding analyses show that the TrxA scaffold can be a major determinant for the binding of peptide aptamers. In addition, we demonstrate that TrxA can correctly display peptide sequences that correspond to the binding domains of natural interaction partners. Therefore, sequence analyses of TrxA-based peptide aptamers, isolated by two-hybrid screening from randomized expression libraries, should also be useful to find cellular binding partners for a given target protein, by homology. Received 1 August 2002; received after revision 17 September 2002; accepted 19 September 2002 RID="*" ID="*"Corresponding author.     

重组类人胶原蛋白Ⅰ的层析分离

目的研究重组类人胶原蛋白的基因工程下游工艺对阴离子交换树脂（DE52）和阳离子交换树脂（CM52）的吸附效果。方法将基因工程菌E．coil BL 21经超声破碎、硫酸铵盐析、超滤浓缩后，再进行批量层析进一步分离纯化。结果得到最佳实验条件：采用阴离子交换树脂，pH值为6．0。盐浓度为0．2mol／L，蛋白进样浓度为0．4％（质量分数）。结论在HCBI的纯化过程中，阴离子交换树脂（DE52）较阳离子交换树脂（CM52）的分离效果更优，最终产物的纯度达到95．98％，回收率为91．7％，纯化倍数为5．3。纯化的类人胶原蛋白Ⅰ经SDS—PAGE电泳测定为电泳纯，分子质量为90ku。     

禽白血病研究进展

禽白血病(AL)是由病毒引起的一种肿瘤性传染病,感染率高,发病率低,是危害养鸡业的主要疾病之一[1].本文主要介绍了其病原学、流行病学、诊断、预防等几个方面的研究近况,旨在加强对禽白血病的防制.     

抵御蠕虫攻击的一种方法-Honeypot系统

Internet大面积遭受蠕虫攻击的事件时有发生，针对这种问题，引入Honeypot技术，结合入侵检测系统(IDS)、数据挖掘提出了一种解决办法：将Honeypot置于DMZ中，利用其欺骗地址空间技术覆盖服务器中没有用到的IP地址，捕获蠕虫；IDS监控流入网络的数据包，对入侵作出反映；系统日志异地保存。该系统能有效抵御目前已经出现的蠕虫攻击，同时对新出现的目前未知的蠕虫攻击也有很好的防御效果。     

Progress in non-viral gene delivery systems fabricated via supramolecular assembly

Gene delivery systems are one of key issues that limit the development of gene therapy. The novel non-viral gene delivery systems fabricated via supramolecular assembly have begun to show increasing promising and applications in gene therapy due to its suitable nanometric size,controllable structure and excellent biocompatibility. In this review, the fundamental and recent progress of non-viral gene supramolecular assembly is reviewed. Artificial viruses the future direction of non-viral gene delivery systems are also described.     

Two virus-encoded RNA silencing suppressors, P14 of Beet necrotic yellow vein virus and S6 of Rice black streak dwarf virus

Functional analysis for gene silencing suppressor of P14 gene of Beet necrotic yellow vein virus and S6 gene of Rice black streak dwarf virus was carried out by agro- infiltration with recombinant vectors of Potato virus X. The phenotype observation of green fluorescent protein (GFP)expression and Northern blot showed that the gene silencing of gfp transgenic Nicotiana benthamiana induced by homologous sequence was strongly suppressed by the immixture infiltration of either the P14 or the $6. In the suppressed plants, the gfp mRNA accumulation was higher than that in the non-suppressed controls and the symptoms caused by PVX infection became more severe, especially the gfp DNA methylation of plant genome was significantly inhabited when co-infiltrated with RBSDV S6 gene. These results suggested that these two virus genes were potentially to encode for proteins as RNA silencing suppressors.     

对虾白斑综合征病毒(WSSV)基因组细菌双杂交系统的构建

细菌双杂交系统是新近建立的一种研究蛋白质间相互作用的方法.应用细菌双杂交系统,分别以pBT、pTRG诱饵质粒构建编码WSSV基因组DNA随机片段的融合表达质粒pBT-wssv、pTRG-wssv.重组质粒共转化双杂交报告菌株XLl-Blue MRF′,通过LB-TCK平板筛选对虾白斑综合征病毒中有相互作用的蛋白质.本研究中我们构建了WSSV基因组细菌双杂交系统,用于筛选WSSV中有相互作用的蛋白,为该病毒功能基因组的研究打下良好的基础.     

重组SARS冠状病毒S蛋白的原核表达研究

目的克隆表达SARS冠状病毒的主要结构蛋白(S蛋白)。方法合成SARS冠状病毒S蛋白特异性基因片断并克隆入pET32a原核表达载体,转化BL21菌,经IPTG诱导高效表达得到重组S蛋白,并通过W estern印迹对重组蛋白质进行鉴定。结果重组蛋白质经镍柱亲和层析得到了部分纯化,免疫动物后得到SARS病毒的多克隆抗体。结论经E lisa检测,表达的重组S蛋白基本具备检测病人血清中抗SARS病毒IgG和IgM的能力,可进一步用于S蛋白功能研究与SARS诊断试剂盒的研制。     

类人胶原蛋白工程菌质粒稳定性研究

目的研究基因重组大肠杆菌补料分批发酵生产类人胶原蛋白过程中质粒稳定性。方法通过平板复制法,观察质粒稳定性在不同生长周期、卡那霉素浓度和不同比生长速率中的变化。结果质粒稳定性受生长周期、卡那霉素和比生长速率的影响。在μ为0.15、卡那霉素浓度为0.005g/L的情况下,工程菌生产效率最理想。结论通过控制发酵过程中的质粒稳定性可以有效地提高菌体产量和目标蛋白产量。     

基于贝叶斯理论的未知病毒检测算法的实现

计算机病毒严重地危害了计算机世界的安全,如何有效地利用已知病毒的特征对未知病毒进行检测是一个非常重要的研究课题。对基于贝叶斯分类理论的未知病毒检测算法给予了系统设计和实现。实验结果表明,这种新的算法对未知病毒具有较高的分辨率。