Insertional re-activation of a chloramphenicol acetyltransferase misfolding mutant protein

The deletion of nine residues from the C-terminus of the bacterialchloramphenicol acetyltransferase (CAT) results in depositionof the mutant protein in cytoplasmic inclusion bodies and lossof chloramphenicol resistance in Escherichia coli. This foldingdefect is relieved by C-terminal fusion of the polypeptide withas few as two residues. Based on these observations, efficientpositive selection for the cloning of DNA fragments has beendemonstrated. The cloning vector encodes a C-terminally truncatedCAT protein. Restriction sites in front of the stop codon allowthe insertion of target DNA, resulting in the production ofproperly folded CAT fusion proteins and regained chloramphenicolresistance. The positive selection of recombinants is accomplishedby growth of transformants on chloramphenicol-containing agarplates. The method appears particularly convenient for the cloningof DNA fragments amplified by the PCR because minimal informationto restore CAT folding can be included in the primers. The cloningof random sequences shows that the folding defect can be relievedby fusion to a wide variety of peptides, providing great flexibilityto the positive selection system. This vector may also contributeto the determination of the role of the C-terminus in CAT folding.     

CAD系统中产品设计三维实体建模方法的研究

三维建模是应用CAD系统进行产品设计的重要步骤。现在市场上已有不少的CAD系统，但是不同系统中的三维建模方法却是基本相同的。在这些三维建模的方法中，往往对用户施加了许多的限制，使得用户无法建立复杂的三维实体。笔者提出了一种新的三维建模方法－折叠法，应用这种方法的概念和原理，可以开发出先进的三维建模CAD系统。折叠二维平面图形构成三维实体是作者提出的一种新概念。     

36 m 高砖砌水塔定向折叠爆破拆除

介绍了复杂环境中砖砌高大水塔的定向折叠爆破拆除.根据周围的环境情况,在施工中采用了开定向窗、挖定位坑等技术措施,确保爆破成功,达到了设计效果.     

Effect of temperature on the development of the yam moth, Dasyses rugosella Stainton (Lepidoptera: Tineidae)

Studies on the effect of temperature on the development of the yam moth, Dasyses rugosella Stainton were carried out in the laboratory at four different temperatures, 20°C, 24°C, 29°C and 33°C. The mean fecundity per female at 20°C, 24°C, 29°C and 33°C was 37.6±1.9, 84.2±2.7, 112.3±3.9 and 109.3±3.9 eggs, respectively. Oviposition period ranged from 4.7 days at 20°C to 2.9 days at 33°C. Hatchability of eggs was highest at 29°C with 70.0% and lowest at 20°C with 15.0%. The developmental time decreased with increase in temperature. The mean developmental time at 20°C, 24°C, 29°C and 33°C was 13.8±0.3, 7.8±0.2, 5.1±0.2 and 3.9±0.2 days for the egg, 90.0±0.7, 50.1±0.7, 36.1±0.8 and 23.6±0.8 days for the larval stages, 22.3±2.6, 12.2±0.2, 10.1±0.1, and 7.8±0.2 days for the pupa and 126.1±0.8, 70.1±0.8, 51.3±0.9, and 35.1±0.8 days for the period from egg hatching to adult emergence, respectively. Since fecundity, oviposition and development of the moth were impaired at 20°C, storage of yam tubers at low temperatures (well below 20°C but not lower than 12°C to avoid chilling injury) will significantly retard the development of D. rugosella and help in its control. Unmated individuals lived longer than their mated counterparts. Adult females were always larger than the males. The wing span of the female ranges from 16.9-18.5 and in males from 13.0-15.0 mm. The body length for males ranged from 5.0-6.5 and was 6.5-9.0 mm for females.     

Designing amino acid sequences to fold with good hydrophobic cores

We present two methods for designing amino acid sequences ofproteins that will fold to have good hydrophobic cores. Giventhe coordinates of the desired target protein or polymer structure,the methods generate sequences of hydrophobic (H) and polar(P) monomers that are intended to fold to these structures.One method designs hydrophobic inside, polar outside; the otherminimizes an energy function in a sequence evolution process.The sequences generated by these methods agree at the levelof 60–80% of the sequence positions in 20 proteins inthe Protein Data Bank. A major challenge in protein design isto create sequences that can fold uniquely, i.e. to a singleconformation rather than to many. While an earlier lattice-basedsequence evolution method was shown not to design unique folders,our method generates unique folders in lattice model tests.These methods may also be useful in designing other types offoldable polymer not based on amino acids     

Alcohol-induced changes of {beta}-lactoglobulin - retinol-binding stoichiometry

It has been demonstrated using CD that ethanol induces importantsecondary structure changes of ß-lactoglobulin. CDspectra indicate that ß-lactoglobulin secondary structure,which is mainly composed of ß-strands, becomes mostly-helical under the influence of the solvent polarity changes.The midpoint of ß-strand/-helix transition in ß-lactoglobulinis observed at dielectric constant {small tilde}60 (35% ethanol;v/v). According to CD measurements, the ethanol-dependent secondarystructure changes are reversible. The alkylation of lysines-NH₂ in ß-lactoglobulin weakens the central ß-barrelstructure, since the ß-strand/-helix transition midpointof alkylated ß-lactoglobulin is shifted to lower ethanolconcentration (25% ethanol; v/v). ß-Lactoglobulinstructural changes are triggering the dissociation of the ß-lactoglobulin- retinol complex as judged from complete quenching of its fluorescencein ethanol concentration >30% (v/v). However, in 20% ethanol(v/v), ß-lactoglobulin still retains most of its nativesecondary structure as shown by CD and, in this condition, oneß-lactoglobulin molecule binds an additional secondretinol molecule. This suggests that the highly populated speciesobserved around 20% ethanol (v/v) might represent an intermediatestate able to bind two molecules of retinol.     

The role of heat-shock and chaperone proteins in protein folding: possible molecular mechanisms

Recently some heat-shock proteins have been linked to functionsof ‘chaperoning’ protein folding in vivo. Here currentexperimental evidence is reviewed and possible requirementsfor such an activity are discussed. It is proposed that onemode of chaperone action is to actively unfold misfolded orbadly aggregated proteins to a conformation from whkh they couldrefold spontaneously; that improperly folded proteins are recognizedby excessive stretches of solvent-exposed backbone, rather thanby exposed hydrophobic patches; and that the molecular mechanismfor unfolding is either repeated binding and dissociation (‘plucking’)or translocation of the protein backbone through a binding cleft(‘threading’), allowing the threaded chain to refoldspontaneously. The observed hydrolysis of ATP would providethe energy for active unfolding. These hypotheses can be appliedto both monomeric folding and oligomeric assembly and are sufficientlydetailed to be open to directed experimental verification.     

广西老堡铅锌矿褶皱形成机制及其控矿规律

老堡铅锌矿床主要赋存在陡山沱组白云岩或老堡组硅质岩中,顺层产出,受地层与褶皱的双重控制。矿区主干褶皱为一复式向斜,由2个次级向斜和1个次级背斜组成。从构造几何学、运动学角度,运用现代测试技术,对矿区的小构造、显微构造和组构进行构造分析后揭示,老堡褶皱构造的形成机制是以弯滑作用为主、兼具弯流作用的纵弯褶皱作用的产物。铅锌矿体主要赋存在褶皱的转折端虚脱部位及两翼滑移量较大的层间滑动破碎带中。     

Bromodomain-like折叠类型模板的设计

针对折叠类型分类中所选天然模板普适性不足的问题,提出了Bromodomain-like折叠类型模板的设计方法.选SCOPe Astral 2.03序列相似度小于40%并且分辨率高于0.25 nm的52个可用Bromodomain-like折叠样本,基于多结构比对结果及数据分析,建立了折叠类型家族模板的设计方法.利用系统聚类方法构建了家族模板的系统聚类图,提出了蛋白质折叠类型模板的设计方法,并用于该折叠类型的模板设计.结果表明：设计模板具有普适性,可用于蛋白质折叠类型分类.     

蛋白质折叠的计算机模拟研究进展

蛋白质折叠是结构生物学领域有待解决的重要科学问题,计算机模拟方法是研究蛋白质折叠的主要手段之一.对蛋白质折叠的计算机模拟研究进展进行了简要介绍,主要内容包括:全原子分子动力学模拟、Gō模型以及弹性网络模型的主要思路及其在蛋白质折叠研究中的应用.