Expression screen by enzyme-linked immunofiltration assay designed for high-throughput purification of affinity-tagged proteins

High-throughput purification of affinity-tagged fusion proteins is currently one of the fastest developing areas of molecular proteomics. A prerequisite for success in protein purification is sufficient soluble protein expression of the target protein in a heterologous host. Hence, a fast and quantitative evaluation of the soluble-protein levels in an expression system is one of the key steps in the entire process. Here we describe a high-throughput expression screen for affinity-tagged fusion proteins based on an enzyme linked immunofiltration assay (ELIFA). An aliquot of a crude Escherichia coli extract containing the analyte, an affinity-tagged protein, is adsorbed onto the membrane. Subsequent binding of specific antibodies followed by binding of a secondary antibody horseradish peroxidase (HRP) complex then allows quantitative evaluation of the analyte using tetramethylbenzidine as the substrate for HRP. The method is accurate and quantitative, as shown by comparison with results from western blotting and an enzymatic glutathione S-transferase (GST) assay. Furthermore, it is a far more rapid assay and less cumbersome than western blotting, lending itself more readily to high-throughput analysis. It can be used at the expression level (cell lysates) or during the subsequent purification steps to monitor yield of specific protein.     

不同生态稻区覆膜旱作稻氮营养生理及抗逆生理特性探讨

不同生态稻区生态环境和栽培管理方式不同,覆膜旱作稻氮营养生理和抗逆生理与常规水作稻有些异同,尤其是体内NO₃^--N和NH₄⁺-N含量及分布差异很大.杭嘉湖平原区覆膜旱作稻生长前期受干热天气影响,其分蘖期、拔节期、孕穗期体内NO₃^--N含量较水作稻有不同程度的降低,而NH₄⁺-N含量则极显著提高;金衢盆地覆膜旱作稻孕穗期体内NO₃^--N含量比常规水作稻高,根部NH₄⁺-N含量则显著降低,茎基部和叶片NH₄⁺-N含量有一定增加.正常气候条件下两生态稻区覆膜旱作稻孕穗期叶片硝酸还原酶(NR)活性和谷氨酰胺合成酶(GS)活性较水作稻均有不同程度的提高;丙二醛(MDA)、可溶性糖及脯氨酸(Pro)含量与水作稻相差不大,表明不同生态稻区的覆膜旱作稻可以通过不同的栽培管理模式,促进生长发育,达到高产目的.     

40种常见花粉相对营养价值评价

本文统计了４０种常见花粉的蛋白质、钙、铁、锌、硒、视黄醇当量、维生素Ｅ和维生素Ｃ的含量并转化为其相对营养价进行分析比较。其相对营养价分别为２８．２２、１６．４３、３０８．６１、３５．０５、４９．１１、１６９６．６０、４１０３．３３和６０．６５。表明花粉营养丰富,视黄醇当量、维生素Ｅ和铁的含量尤其突出,是理想的天然保健食品。     

秦岭汉江流域新发现旧石器遗址的地层与释光年代

汉江流域旧石器时代文化遗存丰富,南方砾石石器工业和北方小石片石器工业类型的遗存并存,对研究中国南北旧石器文化交流和发展演化具有重要的意义。2016-2018年夏天,我们在汉江流域进行野外调查,新发现十个含更新世晚期旧石器遗存的地点,利用热转移光释光（TT-OSL）方法对各遗址的地层进行了测年。测年结果表明,这几处旧石器地点的古人类活动主要发生在距今20-5万年之间,涵盖了L₂、S₁和L₁三个阶段,与之前汉江流域第二级阶地发现的旧石器遗址年代基本一致。新发现的十个旧石器地点丰富了汉水流域旧石器遗存,为进一步研究该区域古人类旧石器遗址年代学和石器技术提供了新的材料。