Pentamerization of single-domain antibodies from phage libraries: a novel strategy for the rapid generation of high-avidity antibody reagents

We describe a novel type of molecule in which single-domain antibodies (sdAbs) isolated from a nai;ve llama single domain antibody library are linked to an oligomerization domain to generate high-avidity, antigen-binding reagents. An sdAb is fused to the B-subunit of Escherichia coli verotoxin, or shiga-like toxin, which self-assembles to form a homopentamer and results in simultaneous sdAb pentamerization and introduction of avidity. Molecular modeling indicated that this fusion protein (PDB: 1OJF), termed pentabody, has structural flexibility for binding to surface-presented antigen. In the instance of an sdAb specific for a peptide antigen, pentamerization resulted in a dramatic increase in functional affinity for immobilized antigen. The pentabody was expressed in high yield in E.coli in a non-aggregated state, and exhibited excellent thermostability and protease resistance. This technology provides a relatively rapid means of generating novel antigen-binding molecules that bind strongly to immobilized antigen. It is expected that pentavalent sdAbs will have general applicability in proteomics, immunochemical staining, cancer diagnosis and other applications in which antigens are presented multivalently.     

人松弛素原样蛋白H2在大肠杆菌中的表达、分离纯化及鉴定

利用聚合酶链式反应方法 ,从含有人前松弛素原cDNA基因的质粒 pMALp2X-hRLXH2上得到人松弛素原H2的编码基因 ,亚克隆到温度诱导型原核表达载体pBV2 2 0上 ,转化大肠杆菌DH5α细胞。经 4 2℃温度诱导 ,获得了目的基因的高效表达。目的蛋白质以包含体的形式存在于大肠杆菌细胞中。菌体经过超声波破碎 ,包含体裂解 ,蛋白质的体外变性还原 ,复性 ,SephadexG 75凝胶过滤层析 ,反相快速蛋白质液相层析等一系列分离纯化过程 ,得到了高纯度的重组Met 人松弛素原样蛋白H2 ,得率约为 2～ 3mg/L。对纯化的目的蛋白质进行了SDS PAGE电泳纯度分析 ,氨基酸组成分析 ,通过基质辅助激光解吸电离飞行时间质谱法测得目的蛋白质的分子量为 183 90 .4 (理论计算值为 183 92 .3 )。     

Toward Long‐Term Stable and Highly Efficient Perovskite Solar Cells via Effective Charge Transporting Materials

Perovskite solar cells (PSCs) have advanced quickly with their power conversion efficiency approaching the record of silicon solar cells. However, there is still a big challenge to obtain both high efficiency and long‐term stability for future commercialization of PSCs. The major instability issue is associated with the decomposition or phase transition of perovskite materials that are believed to be intrinsically unstable under outdoor working conditions. Herein, the authors review the approaches that marked important progress in developing new functional electron/hole transporting materials that enabled highly efficient and stable PSCs. The findings that accelerate charge diffusion and that suppress the irrevocable loss of ions diffusing out of perovskite materials and other diffusion processes are highlighted. In addition, derivative interface engineering methods to control the diffusion process of charges/ions/molecules are also reviewed. Finally, the authors propose key research issues in charge transporting materials and interface engineering with regard to the important diffusion processes that will be one of the keys to realize highly efficient and long‐term stable PSCs.     

A Systems Approach to Sustainable Technical Product Design

Many existing methods for sustainable technical product design focus on environmental efficiency while lacking a framework for a holistic, sustainable design approach that includes combined social, technical, economic, and environmental aspects in the whole product life cycle, and that provides guidance on a technical product development level. This research proposes a framework for sustainable technical product design in the case of skis. We developed a ski under the Grown brand, benchmarked according to social, environmental, economic, and technical targets, following an initial sustainability assessment, and delivered the first environmental life cycle assessment (ELCA) and the first social life cycle assessment (SLCA) of skis. The framework applies a virtual development process as a combination of ELCA to calculate the environmental footprint as carbon equivalents of all materials and processes and a technical computer‐aided design (CAD) and computer‐aided engineering (CAE) simulation and virtual optimization using parameter studies for the nearly prototype‐free development of the benchmarked skis. The feedback loops between life cycle assessment (LCA) and virtual simulation led to the elimination of highly energy intensive materials, to the pioneering use of basalt fibers in skis, to the optimization of the use of natural materials using protective coatings from natural resins, and to the optimization of the production process. From an environmental perspective, a minimum 32% reduction in carbon equivalent emissions of materials in relation to other comparably performing skis has been achieved, as well as a pioneering step forward toward transparent communication of the environmental performance by the individual, comparable, and first published ski carbon footprint per volume unit.     

《酶工程》教学与课程建设的探讨

酶工程是生物技术专业的主干课程,其主要内容是研究酶的生产和应用。为了使酶工程的教学适合时代发展的需要,培养合格的从事酶工程研究及生产的技术人才,以多年酶工程教学实践为基础,集中介绍该门课程教学与建设的体会及其思考。     

以达托霉素产生菌菌株改造为主线的微生物工程综合实验的探索和实践

为使本科学生更好地掌握微生物工程的基本实验操作技能和前沿研究技术,探索将教师的科研成果转化为教学实验,构建了以达托霉素产生菌菌株改造为主线的微生物工程综合实验。在教学模式、实验内容设计、教学安排和考核方式等方面进行实践,取得了较好的教学效果,培养了本科学生的科研技能、科研思维和研究素质。     

乳酸菌在农业和食品加工中的应用研究进展

乳酸菌是发酵糖类物质的主要终产物为乳酸的细菌总称。常见的乳酸菌包括乳杆菌属(Lactobacillus)、片球菌属(Pediococcus)、明串珠菌属(Leuconostoc)、乳球菌属(Lactococcus)、链球菌属(Streptococcus)和双歧杆菌属(Bifdobacterium)等少数属的种类。许多乳酸菌属于人和动物胃肠道、泌尿生殖道、母乳、口腔等的正常菌群,能产生多种生物活性物质,具有调节肠道菌群平衡、增强机体免疫力、促进机体生长等益生功能,因而乳酸菌具有重要的应用价值。文本综述了乳酸菌在畜禽养殖饲料添加剂、青贮饲料发酵、水产养殖及植物病害防治等领域的应用研究进展,尤其在“饲料禁抗、养殖减抗、产品无抗”的养殖背景下,乳酸菌制剂是替代抗生素的最佳选择。同时也总结了乳酸菌在奶制品、肉制品加工以及植物基饮料、低糖食品和饮料发酵等方面的开发潜力,展望了乳酸菌研究与应用的发展趋势。     

Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold

Genetically engineered rice (Oryza sativa L.) with the ability to synthesize glycinebetaine was established by introducing the codA gene for choline oxidase from the soil bacterium Arthrobacter globiformis. Levels of glycinebetaine were as high as 1 and 5 mol per gram fresh weight of leaves in two types of transgenic plant in which choline oxidase was targeted to the chloroplasts (ChlCOD plants) and to the cytosol (CytCOD plants), respectively. Although treatment with 0.15 m NaCl inhibited the growth of both wild-type and transgenic plants, the transgenic plants began to grow again at the normal rate after a significantly less time than the wild-type plants after elimination of the salt stress. Inactivation of photosynthesis, used as a measure of cellular damage, indicated that ChlCOD plants were more tolerant than CytCOD plants to photoinhibition under salt stress and low-temperature stress. These results indicated that the subcellular compartmentalization of the biosynthesis of glycinebetaine was a critical element in the efficient enhancement of tolerance to stress in the engineered plants.     

基因工程技术在食品品质改良中的应用

概述了用基因工程技术改良食品的食用品质,提高食品加工性能的研究和应用现状。