家蚕核多角体病毒双穿梭载体的构建及GFP和HBeAg融合蛋白的表达

用ＡｃＮＰＶ穿梭载体Ａｃ－Ｂａｃｍｉｄ与野生型ＢｍＮＰＶＤＮＡ共转染家蚕ＢｍＮ细胞,通过同源重组得到整合了ｌａｃＺ／Ｔｎ７ａｔ／ｍｉｎｉＦ表达盒的ＢｍＮＰＶＢａｃｍｉｄ,除能在大肠杆菌／ＢｍＮ细胞中穿梭复制外,还能感染ＢｍＮ－ＰＶ的非允许细胞Ｓｆ９,成功地构建了ＢｍＮＰＶ的双穿梭载体．经与重组转座质粒ｐＦＧＨｅ转座,获得了插入ｇｆｐ／ＨＢＶｅ融合基因的重组ｒＢｍ－Ｂａｃｍｉｄ（ｒＢｍＧＨｅ）,在家蚕细胞中表达了既能发射绿色荧光又具有ＨＢｅＡｇ抗原性的双功能融合蛋白．Ｂｍ－Ｂａｃｍｉｄ为杆状病毒ＢａｃｔｏＢａｃ策略增添了一个新成员     

突变型绿色荧光蛋白基因在昆虫细胞和幼虫中的表达

用细菌—杆状病毒穿梭系统（Ｂａｃ－ｔｏ－Ｂａｃ）将一套含有绿色荧光蛋白基因（ｇｆｐＳ６５Ｔ）和完整多角体蛋白基因的表达盒转座到ＡｃＮＰＶ－Ｂａｃｍｉｄ的Ｔｎ７转座接受位点,从而构建出一种带有ｇｆｐ基因和完整多角体基因的重组杆状病毒,用这一重组病毒感染细胞能稳定地形成多角体,并在紫外光照射下产生强烈的绿色荧光,以重组病毒感染粉纹夜蛾（Ｔｒｉｃｈｏｐｌｕｓｉａｎｉ）幼虫后,在阳光下即可看到发射绿色荧光的幼虫,在手提式长波紫外光照射下绿色荧光更为强烈,带绿色荧光的幼虫在昆虫生态学和杆状病毒流行病学等研究领域有广泛的用途     

B组轮状病毒WH-1株nsp2基因的原核表达与抗体制备

根据B组轮状病毒(GBRV)WH-1株nsp2基因的全序列,设计引物,用PCR的方法扩增得到nsp2基因的编码区.将其片段克隆到原核表达载体pGEX-KG上,经IPTG诱导,在E.coliDH5α菌株中得到高效表达.表达的GST融合蛋白大小为61×103,经SDS-PAGE分离纯化,免疫小白鼠制备了多克隆抗体.抗体经1∶3 000倍稀释后用于Western Blot分析,获得特异性显色信号.所表达的蛋白和制备的抗体可用于蛋白结构和功能的进一步研究.     

海藻酸钠/大豆蛋白共混凝胶微球的结构

利用钙离子交联海藻酸钠／大豆分离蛋白共混溶液，制得海藻酸钠／大豆分离蛋共混凝胶微球．结果表明，海藻酸钠和大豆分离蛋白质量配比的不同以及各组分间相互作用的变化，微球呈现不同的微观结构．将微球干燥后置于水中溶胀，微球的尺寸无法回复到干燥前的尺寸，这是由于真空干燥处理使水分子挥发，促进微球内组分间形成了强的氢键作用所致．此外，用碱处理该共混微球，发现由于大豆分离蛋白溶解以及部分钙离子被置换析出，微球塌陷且内部形成了大孔．     

On the Nonrepresentability of Functions of Several Variables by Quadratures

The paper completes the construction of a multidimensional topological version of differential Galois theory. We construct a rich class of germs of functions of several variables which is closed under superpositions and other natural operations. The main theorem describes the behavior of the monodromy groups of such germs under the natural operations. As a result, we obtain topological obstructions to the representability of functions by quadratures, which give the strongest known statements about unsolvability of equations in closed form.     

Fluorescence anisotropy decay study of self-association of bacterial luciferase intermediates

The fluorescence dynamics parameters of the fluorescent transient flavin-luciferase species from the typesVibrio fischeri andPhotobacterium leiognathi are presented. The fluorescence anisotropy decay is a single exponential function for both types. The correlation time is 70 ns for theP. leiognathi fluorescent transient intermediate (2°C, aqueous buffer, pH 7.0), consistent with the rotational correlation time of the luciferase macromolecule (77 kD) to which the flavin fluorophore is rigidly attached. In contrast, for theV. fischeri species the observed correlation time for the anisotropy decay function is 133 ns. This suggests that protein self-association occurs in theV. fischeri case and this is confirmed by filtration, where the fluorescent transient fromV. fischeri does not pass through a 100,000 molecular weight cutoff membrane, whereas theP. leiognathi species does. The filtration method also demonstrates self-association in the luciferase peroxyflavin and photoflavin fromV. fischeri. A monomer-dimer equilibrium also explains the previously reported high correlation times for theV. harveyi luciferase-flavin species. It is proposed that the self-association competes with the lumazine protein interaction in the bioluminescence reaction.     

纳米孔壁面作用对蛋白质过孔影响的粗粒化分子动力学模拟

基于粗粒化分子动力学方法模拟电驱动蛋白质过孔过程,研究纳米孔-水/纳米孔-蛋白质相互作用对电泳迁移率的影响;用操控式分子动力学模拟分析蛋白质在不同相互作用下过孔摩擦系数和摩擦阻力.研究发现：蛋白质黏附纳米孔壁面对其过孔特性影响并不明显,而纳米孔-水相互作用对蛋白质过孔电泳迁移率和摩擦系数影响较大.随纳米孔-水相互作用增强,纳米孔壁面与蛋白质附近水分子运动差异显现,蛋白质过孔摩擦阻力显著增大,过孔摩擦系数随之增大,进而影响蛋白质过孔电泳迁移率.所得结果可为纳米孔材料设计提供理论指导.     

Classification of recyclables using laser-induced breakdown spectroscopy for waste management

With the ever-increasing amount of generated waste governments around the world are looking for, and implementing, ways to minimize waste output and maximize waste recovery. The main difficulties are sorting waste items, identifying the different types of plastics, and the time taken to sort them manually. Bioplastics such as polylactic acid and Novatein thermoplastic protein can be incorporated into the recycling stream to minimize waste. Laser-induced breakdown spectroscopy spectra analyzed by k-nearest neighbor and soft independent modeling by class analogy were investigated as methods that can rapidly identify recyclables. Raw, peak normalized, and total intensity normalized spectra were used to identify which would improve classification. Laser-induced breakdown spectroscopy spectra were generated by single laser shots to different locations on nine samples, glass (brown, green, and clear), tin, aluminum, polylactic acid, Novatein, polyethylene terephthalate, and high-density polyethylene. To prove that the system has the potential to be used on a waste sorting stream an autofocus unit was developed to move the laser-induced breakdown spectroscopy beam into focus on the different sample geometries. Two classification methods were investigated, soft independent modeling by class analogy and the k-nearest neighbors algorithm. k-Nearest neighbors on raw spectra produced the best results. Discrimination between bioplastics and plastics were 100%. Glass samples could not be reliably distinguished from each other. Surface contamination produced three misclassifications from 450 spectra. Similar results were obtained when the spectral range was reduced from 182.26–908.07?nm to 313.20–495.12?nm.     

单分子荧光检测在生命科学中的应用

文章对单分子荧光检测在分子马达、离子通道、信号分子、蛋白折叠、蛋白构象变化动力学、酶活性反应、细胞过程实时观察等生命科学领域中的应用进行了介绍.这些研究结果表明,单分子荧光检测在研究生物大分子的活动规律与机制方面不但有着无法替代的优越性,而且有着广阔的发展空间.     

Study of 19Na at SPIRAL

The excitation function for the elastic-scattering reaction p( ¹⁸Ne, p) ¹⁸Ne was measured with the first radioactive beam from the SPIRAL facility at the GANIL laboratory and with a solid cryogenic hydrogen target. Several broad resonances have been observed, corresponding to new excited states in the unbound nucleus ¹⁹Na. In addition, two-proton emission events have been identified and are discussed.