Aminoacylation of synthetic DNAs corresponding to Escherichia coli phenylalanine and lysine tRNAs

Synthetic DNA oligomers (tDNAs) corresponding to Escherichia coli tRNA(Phe) or tRNA(Lys) have been synthesized with either deoxythymidine (dT) or deoxyuridine (dU) substituted in the positions occupied by ribouridine or its derivatives. The tDNAs inhibited the aminoacylation of their respective tRNAs with their cognate amino acids, but not the aminoacylation of tRNA(Leu) with Leu. In the presence of aminoacyl-tRNA synthetase, species of both a tDNA(Phe) synthesized with a 3' terminal riboadenosine and a tDNA(Lys) containing only deoxynucleotides could be aminoacylated with the appropriate amino acids, although the Michaelis constant Km and observed maximal rate Vmax values for aminoacylation were increased by three- to fourfold and decreased by two- to threefold, respectively. The aminoacylation of synthetic tDNAs demonstrates that the ribose backbone of a tRNA is not absolutely required for tRNA aminoacylation.     

End-to-end stacking and liquid crystal condensation of 6 to 20 base pair DNA duplexes

Short complementary B-form DNA oligomers, 6 to 20 base pairs in length, are found to exhibit nematic and columnar liquid crystal phases, even though such duplexes lack the shape anisotropy required for liquid crystal ordering. Structural study shows that these phases are produced by the end-to-end adhesion and consequent stacking of the duplex oligomers into polydisperse anisotropic rod-shaped aggregates, which can order into liquid crystals. Upon cooling mixed solutions of short DNA oligomers, in which only a small fraction of the DNA present is complementary, the duplex-forming oligomers phase-separate into liquid crystal droplets, leaving the unpaired single strands in isotropic solution. In a chemical environment where oligomer ligation is possible, such ordering and condensation would provide an autocatalytic link whereby complementarity promotes the extended polymerization of complementary oligomers.     

Optical detection of DNA conformational polymorphism on single-walled carbon nanotubes

The transition of DNA secondary structure from an analogous B to Z conformation modulates the dielectric environment of the single-walled carbon nanotube (SWNT) around which it is adsorbed. The SWNT band-gap fluorescence undergoes a red shift when an encapsulating 30-nucleotide oligomer is exposed to counter ions that screen the charged backbone. The transition is thermodynamically identical for DNA on and off the nanotube, except that the propagation length of the former is shorter by five-sixths. The magnitude of the energy shift is described by using an effective medium model and the DNA geometry on the nanotube sidewall. We demonstrate the detection of the B-Z change in whole blood, tissue, and from within living mammalian cells.     

Single-molecule cut-and-paste surface assembly

We introduce a method for the bottom-up assembly of biomolecular structures that combines the precision of the atomic force microscope (AFM) with the selectivity of DNA hybridization. Functional units coupled to DNA oligomers were picked up from a depot area by means of a complementary DNA strand bound to an AFM tip. These units were transferred to and deposited on a target area to create basic geometrical structures, assembled from units with different functions. Each of these cut-and-paste events was characterized by single-molecule force spectroscopy and single-molecule fluorescence microscopy. Transport and deposition of more than 5000 units were achieved, with less than 10% loss in transfer efficiency.     

Structure and flexibility adaptation in nonspecific and specific protein-DNA complexes

Interaction of regulatory DNA binding proteins with their target sites is usually preceded by binding to nonspecific DNA. This speeds up the search for the target site by several orders of magnitude. We report the solution structure and dynamics of the complex of a dimeric lac repressor DNA binding domain with nonspecific DNA. The same set of residues can switch roles from a purely electrostatic interaction with the DNA backbone in the nonspecific complex to a highly specific binding mode with the base pairs of the cognate operator sequence. The protein-DNA interface of the nonspecific complex is flexible on biologically relevant time scales that may assist in the rapid and efficient finding of the target site.     

The crystal structure of TAL effector PthXo1 bound to its DNA target

DNA recognition by TAL effectors is mediated by tandem repeats, each 33 to 35 residues in length, that specify nucleotides via unique repeat-variable diresidues (RVDs). The crystal structure of PthXo1 bound to its DNA target was determined by high-throughput computational structure prediction and validated by heavy-atom derivatization. Each repeat forms a left-handed, two-helix bundle that presents an RVD-containing loop to the DNA. The repeats self-associate to form a right-handed superhelix wrapped around the DNA major groove. The first RVD residue forms a stabilizing contact with the protein backbone, while the second makes a base-specific contact to the DNA sense strand. Two degenerate amino-terminal repeats also interact with the DNA. Containing several RVDs and noncanonical associations, the structure illustrates the basis of TAL effector-DNA recognition.     

Charge migration in DNA: ion-gated transport

Electron hole (radical cation) migration in DNA, where the quantum transport of an injected charge is gated in a correlated manner by the thermal motions of the hydrated counterions, is described here. Classical molecular dynamics simulations in conjunction with large-scale first-principles electronic structure calculations reveal that different counterion configurations lead to formation of states characterized by varying spatial distributions and degrees of charge localization. Stochastic dynamic fluctuations between such ionic configurations can induce correlated changes in the spatial distribution of the hole, with concomitant transport along the DNA double helix. Comparative ultraviolet light-induced cleavage experiments on native B DNA oligomers and on ones modified to contain counterion (Na(+))-starved bridges between damage-susceptible hole-trapping sites called GG steps show in the latter a reduction in damage at the distal step. This reduction indicates a reduced mobility of the hole across the modified bridge as predicted theoretically.     

竹纤维增强聚酰胺树脂复合材料界面改性

对竹纤维增强聚酰胺树脂复合材料界面改性剂及其界面改性机理进行研究,以聚乙二醇和马来酸酐为原料,用热催化法合成具有线型结构的羧化聚醚。通过红外光谱分析表明,在聚醚链上成功地接枝马来酸酐;经羧化聚醚界面改性剂改性后,羧化聚醚中的马来酸酐可与竹纤维中的羟基发生酯化反应;聚酰胺树脂端氨基和酰胺键中的亚氨基与羧化聚醚中马来酸酐发生酰胺化反应。经2%羧化聚醚改性后,竹纤维聚酰胺树脂复合材料强度性能指标和热变形温度均有大幅度地提高,竹纤维增强效果显著。     

Deoxycytidylate and deoxyguanylate kinase activity in pneumococci after exposure to known polyribonucleotides

Polycytidylic acid and to a lesser extent polyadenylic acid enhance the activity of deoxycytidylate and deoxyguanylate kinases in resting cell suspensions of encapsulated pneumococci. The active intracellular materials appear to be oligomers of A and C, respectively. The stimulation of the kinase activities is amino-acid dependent and can be abolished by the addition of chloramphenicol. The addition of all eight naturally occurring deoxyribonucleosides and deoxyribonucleotides to cell suspensions containing the homopolymers leads to a selective enhancement of DNA synthesis.     

聚酰胺分离纯化翅果油树叶黄酮工艺研究

采用聚酰胺树脂分离纯化翅果油树(Elaeagnus mollis Diels)叶黄酮,通过不同条件下两种聚酰胺树脂(100-200目、60-100目)对翅果油树叶黄酮静态和动态吸附与解吸特性的研究,确定最佳精制工艺。结果表明,100-200目聚酰胺树脂对翅果油树叶黄酮的吸附性能较好,饱和吸附量为0.153mg/g,解吸率为85.88%;最佳工艺条件为吸附液pH 4.0～6.0、解吸液体积分数为60%、V(吸附液)∶m(树脂)=6∶1、洗脱剂用量为1.5BV(柱体积);在此条件下,聚酰胺树脂可重复使用4～5次。     

Saturation of cubic optical nonlinearity in long-chain polyene oligomers

The scaling of the cubic nonlinearity gamma with chain length in polyenic molecules has received considerable theoretical attention. Earlier experimental investigations have been restricted to oligomers with fewer than 20 double bonds because of problems associated with the synthesis and solubility of conjugated molecules. These synthetic difficulties have been overcome in the present study by the use of modern living polymerization techniques. Solution measurements of gamma as a function of chain length in long-chain (up to 240 double bonds) model polyene oligomers are reported. A saturation of the increase of gamma with chain length is observed, and the onset of this saturation occurs for chain lengths considerably longer than predicted from theory.     

丹参地上部分正丁醇萃取部位化学成分检测及其抗脑缺血研究

筛选丹参地上部分抗脑缺血的活性部位,并检测其所含化学成分,对丹参的开发和利用具有重要意义。采用D101大孔吸附树脂吸附丹参地上部分正丁醇萃取部位,将其分离纯化为多个部位。以昆明小鼠常压缺氧模型和不完全性脑缺血模型筛选有效部位;将筛选出的活性部位再用聚酰胺柱分离为不同部位,选用上述模型进行活性部位筛选;并用高效液相检测筛选的活性部位所含化学成分。结果表明:D-101大孔树脂经体积分数为50%和75%乙醇洗脱部位能显著增加小鼠常压耐缺氧时间,延长小鼠全脑缺血后的存活时间(P0.01);聚酰胺柱分离后的水洗部位可显著增加小鼠常压耐缺氧时间和小鼠全脑缺血后的存活时间(P0.01);高效液相检测聚酰胺柱分离的水洗部位含有原儿茶醛、咖啡酸、丹参素。丹参地上部分正丁醇萃取部位聚酰胺柱分离的水洗部位为有效部位,检测出有效成分为原儿茶醛、咖啡酸、丹参素。     

Second structural motif for recognition of DNA by oligonucleotide-directed triple-helix formation

Relative orientations of the DNA strands within a purine.purine.pyrimidine triple helix have been determined by affinity cleaving. A purine-rich oligonucleotide bound in the major groove of double-helical DNA antiparallel to the Watson-Crick purine strand. Binding depended upon the concentration of multivalent cations such as spermine or Mg2+, and appeared to be relatively independent of pH. Two models with specific hydrogen-bonding patterns for base triplets (G.GC, A.AT, and T.AT) are proposed to explain the sequence specificity of binding. The two models differ in the conformation about the glycosyl bond (syn or anti) and the location of the phosphate-deoxyribose backbone in the major groove of DNA. This motif broadens the structural frameworks available as a basis for the design of sequence-specific DNA binding molecules.     

桑葚花色素甙的提纯和分析

为了探讨用于分析有色植物的花色素甙成分的简易迅捷方法,采用酸性乙醇浸提桑葚中的花色素甙,以黑米的花色素甙提取物为参照标准,选用聚酰胺薄膜圆盘层析对提取物进行定性定量分析.桑葚花色素甙的主要成分为矢车菊花色素-3-O-葡萄糖甙.聚酰胺薄膜圆盘层析测定有色植物中的花色素甙含量简便、快捷、准确、可靠,适用于花色素甙的初步鉴定,并可用于野外分析.     

Common structure of soluble amyloid oligomers implies common mechanism of pathogenesis

Soluble oligomers are common to most amyloids and may represent the primary toxic species of amyloids, like the Abeta peptide in Alzheimer's disease (AD). Here we show that all of the soluble oligomers tested display a common conformation-dependent structure that is unique to soluble oligomers regardless of sequence. The in vitro toxicity of soluble oligomers is inhibited by oligomer-specific antibody. Soluble oligomers have a unique distribution in human AD brain that is distinct from fibrillar amyloid. These results indicate that different types of soluble amyloid oligomers have a common structure and suggest they share a common mechanism of toxicity.     

环介导等温扩增技术的研究进展

环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术是利用2对特殊设计的引物和一种具有链置换活性的DNA聚合酶,在恒温条件下特异、高效地扩增DNA的新技术,扩增产物为一系列反向重复的靶序列构成的茎环结构和多环花椰菜样结构的DNA片段混合物,电泳图谱呈阶梯式带状分布,反应伴有白色的副产物焦磷酸镁沉淀产生,可通过肉眼定性判断反应结果.LAMP技术已在核酸的科学研究、疾病诊断和动物胚胎性别的鉴定等领域得到了广泛的应用.从LAMP技术的原理、引物组成及反应过程、技术特点、方法延伸和应用等方面对其进行了概述,以期为同类研究提供参考.     

金银花中绿原酸的提取纯化方法研究

[目的]对金银花中绿原酸的提取和纯化方法进行研究。[方法]用浓度70％乙醇和水做预试验确定提取溶剂,然后通过正交试验考察金银花中绿原酸提取工艺的最佳条件。在纯化试验中选用乙酸乙酯萃取法和聚酰胺层析法,得出纯化的最佳方案,并对聚酰胺层析的层析条件和不同洗脱剂的洗脱效果进行考察。[结果]试验表明,金银花中绿原酸的最佳提取条件为：以浓度70％的乙醇作提取溶液,加醇量为12倍,提取3次,每次2h。平均提取率为6．62％。乙酸乙酯萃取法纯化后绿原酸纯度为18．4％;吸附有绿原酸的聚酰胺以浓度30％的乙醇洗脱效果较好,得到的绿原酸纯度为33．2％。[结论]采用聚酰胺层析较有机溶剂纯化法操作简便、纯度高、无有机溶剂残留且成本低。