氧化硫硫杆菌重组质粒pSDT125的构建及其在大肠杆菌之间的转移

对氧化硫硫杆菌(Thiobacillus thiooxidans)隐蔽性质粒pTt12进行了限制性酶切分析,并绘出了限制性酶切图谱,其大小为13.7Kb.将pTt12质粒与pBR325质粒重组,得到了重组质粒pSDT125(Cm',Ap',Tc~s).氧化硫硫杆菌重组质粒pSDT125本身不具有接合转移能力,但它在广泛寄主范围的转移性质粒RP4的带动下以较高的频率在大肠杆菌(Escherichia coli)之间转移,表明在氧化硫硫杆菌质粒pTt12上存在着与接合转移有关的DNA序列.     

茶刺蛾[Darna trima(more)]颗粒体病毒鉴定及核酸与蛋白质特性的初步研究

1985年从四川珙县分离到的茶刺蛾病原物,经感染试验,组织病理研究,包涵体及病毒粒子超微结构研究,核酸类型鉴别等,定名该病原物为茶刺蛾颗粒体病毒.另对该病毒核酸作了限制性内切酶分析和热变性试验,测得该病毒核酸为双链DNA分子,分子量为64.11×10~6d,93.87kb,Tm值为67.3℃,(G+C)含量为32.7％,包涵体蛋白的氨基酸组分中Asp、Glu和Arg含量高,占氨基酸总量的34.12％,His,Cys和Met含量很少。该病毒对茶刺蛾幼虫有很强的感染力,氨基酸在生物防治上有潜在应用前景.     

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。