黑胸大蠊（Periplaneta fuliginosa）病毒的分离及某些特性

从黑胸大蠊(Periplaneta fuliginosa)自然罹病的虫尸中分离得到一株非包涵体病毒。将病毒悬液均匀拌入无菌饲料并供食154～169日龄黑胸大蠊健康若虫时,能使其感染、发病,死亡率可达98％以上。在电子显微镜下观察时,病毒为球形二十面体颗粒,直径约23nm。病毒悬液具有典型核蛋白紫外吸收光谱。病毒用DNase和RNase处理并经吖啶橙染色、二苯胺和苔黑酚试验及甲醛反应证明:该病毒含有单链DNA。以上特性与细小病毒科的特征有点类似。     

兔出血症病毒核酸的某些理化性质的研究

本文对我国无锡分离的兔出血症病毒A_2R-3毒株核酸的某些理化性质进行了研究。采用孚尔根染色、二苯胺反应和核酸酶解实验证实病毒核酸为DNA类型。吖啶橙染色、甲醛反应、核酸酶S_1消化和核酸热变性实验表明病毒核酸为单链型。核酸电泳呈单一组分。电境观察显示核酸分子链呈线状,平均长度约为2.15μ。计算分子量约为2.1—2.5×10~6d。核酸碱基组盛为A25.34、T29.37、G23.85、C21.43、(G C)克分子百分比值为45.28。结合以前的报道、我们认为:兔出血症病毒可以归类于细小病毒科。     

Responses to selection on genotypic or phenotypic values in the presence of genes with major effects

Summary Average genotypic responses were compared after selection for genotypic values and for phenotypic values on the basis of single-gene models and multigene models in simulated livestock populations. Single-gene models dealt with single gene control of the genetic differences between animals, while multigene models considered a collection of genes with various magnitudes of effects on a trait. In each case, selection lasted through discrete generations until the fixation of the gene frequencies occurred. Generations to reach fixation were used to compare various models, and the two criteria for selection, for their efficiency in selection. Implications of using these models versus using infinitesimal models for selection in practice are presented.     

Expression of aChlamydia anticarbohydrate single-chain antibody as a maltose binding fusion protein

A single-chain antibody fragment has been constructed for an antibody that binds to theChlamydia specific carbohydrate structure of the lipopolysaccharide. Single-chain protein was expressed and secreted into the periplasmic space ofE. coli as a fusion protein with the maltose binding protein. The fusion protein was purified in one step by virtue of its ability to bind to maltose. In a sandwich ELISA, the eluted protein boundChlamydia lipopolysaccharide, which demonstrates that the single-chain protein domain will function as part of a fusion protein. The expression of maltose binding fusion proteins into the periplasmic space could be used for production of other single-chain antibodies or protein fragments requiring appropriate folding and disulfide bond formation.     

Growth yields and the efficiency of oxidative phosphorylation of Paracoccus denitrificans during two- (carbon) substrate-limited growth

Paracoccus denitrificans was grown aerobically during two-(carbon)substrate-limitation on mannitol and methanol in chemostat cultures. Theoretical growth parameters were calculated based on the presence of 2 or 3 sites in the electron-transport chain of Paracoccus denitrificans. Experimental growth parameters determined during two-(carbon)substrate growth were conform to the presence of 3 sites of oxidative phosphorylation, while cells grown only on mannitol possessed 2 sites. The maximum growth yield on adenosine triphosphate (ATP), corrected for maintenance requirements, determined in chemostat experiments in which the methanol concentration is less than 2.11 times the mannitol concentration was 8.6 g of biomass. When the methanol concentration was more than 2.11 times the mannitol concentration the maximum growth yield on adenosine triphosphate decreased due to the more energy consuming process of CO₂-assimilation. Cells use methanol only as energy source to increase the amount of mannitol used for assimilation purposes. When the methanol concentration in chemostat experiments was more than 2.11 times the mannitol concentration, all mannitol was used for assimilation and excess energy derived from methanol was used for CO₂-assimilation via the ribulose-bisphosphate cycle. The synthesis of ribulosebisphosphate carboxylase was repressed when the methanol concentration in chemostat experiments was less than 2.11 times the mannitol concentration or when Paracoccus denitrificans was grown in batch culture on both methanol and mannitol. When in chemostat experiments the methanol concentration was more than 2.11 times the mannitol concentration ribulose-bisphosphate carboxylase activity could be demonstrated and CO₂-assimilation will occur. It is proposed that energy produced in excess activates or derepresses the synthesis of the necessary enzymes of the ribulose-bisphosphate cycle in Paracoccus denitrificans. Consequently growth on any substrate will be carbonas well as energy-limited. When methanol is present in the nutrient cells of Paracoccus denitrificans synthesize a CO-binding type of cytochrome c, which is essential for methanol oxidase activity.The reason for the increase in efficiency of oxidative phosphorylation from 2 to 3 sites is most probably the occurrence of this CO-binding type of cytochrome c in which presence electrons preferentially pass through the a-type cytochrome region of the electron-transport chain.Non Standard Abbreviations X prosthetic group of methanol dehydrogenase - q _substrate specific rate of consumption of substrate (mol/g biomass. h.) - Y _substrate, Y _substrate ^MAX are respectively the growth yield and the maximum growth yield corrected for maintenance requirements (g biomass/mol) - m _substrate maintenance requirement (mol substrate/g biomass) - specific growth rate (h^-1) - M [methanol]/[mannitol] ratio in the nutrient - N part of mannitol that is assimilated when M=o - R _m amount of methanol-equivalents that has the same energy content as 1 mannitol-equivalent - P/O _N , P/O _F , P/O _X is the amount of ATP produced during electron-transport of two electrons from respectively NADH+H⁺, FADH₂ and XH₂ to oxygen     

Purification and characterization of a highly acidic 2Fe-ferredoxin from Halobacterium of the dead sea

A 2Fe-ferredoxin from Halobacterium of the Dead Sea has been purified by chromatography on Sepharose and DEAE-cellulose, using decreasing concentration gradients of ammonium sulfate. Its amino acid composition reveals an extremely high excess of acidic amino acid residues: 44 glutamate and aspartate residues (of which 4 are in the amide form), compared to 6 lysines and arginines, as well as a high content of aromatic amino acids. The molecular weight of this ferredoxin was found to be 14,000 by amino acid composition, sedimentation equilibrium, and iron content. The millimolar coefficients at the maxima of the visible absorption spectrum are: 28.0 (277 nm), 12.2 (330 nm), 9.1 (420 nm), and 8.3 (465 nm). The optical properties—absorption and CD spectra in the visible region—of this ferredoxin are very similar to those of plant and algal ferredoxins, whereas its redox potential is much higher: ?345 ± 5 mV (at pH 7.3, 0.5 m NaCl). Although it is reduced by illuminated chloroplasts, it cannot mediate the photoreduction of NADP in their presence. Data reported elsewhere suggest that its physiological function might be to serve as an electron donor for nitrite reduction.     

The utilization of bromodeoxyuridine incorporation into DNA for the analysis of cellular kinetics

Recently developed differential staining techniques based on the incorporation of bromodeoxyuridine (BUdR) into DNA permits the unequivocal identification of metaphase cells which have replicated once, twice, and three or more times. This technique has the potential of being utilized in the examination of kinetics of dividing cell populations. This potential is examined in a phytohemagglutinin-stimulated lymphocyte system. Determinations of the effect of increasing concentrations of BUdR on the distribution of metaphase cells between different generation cycles reveals no inhibition of cellular kinetics below 35 μM. The ability to distinguish third generation metaphase cells from subsequent generations is examined through the determination of “labelled” centromeric regions. The applicability of this system to current cellular kinetics is discussed.     

1/f Membrane noise generated by diffusion processes in unstirred solution layers

A mathematical treatment is given for 1/f noise observed in the ion transport through membranes. It is shown that this noise can be generated by current or voltage fluctuations which occur after step changes of the membrane permeability. Due to diffusion polarization in the unstirred solution layers near the membrane these fluctuations exhibit a 1/t time course which produces noise with a 1/f frequency dependence. The spectral density of 1/f noise is calculated for porous membranes with random switches between a finite and zero pore permeability. A wide frequency range and a magnitude of 1/f noise are obtained which are compatible with experimental data of 1/f noise reported for nerve membranes.Supported by Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 38 Membranforschung.     

The physico-chemical mechanism of mediated transport. I. Facilitated diffusion

On the basis of the currently accepted model for the cell membrane structure, a physico-chemical model for mediated transport is developed and solved for the case of polar non-electrolyte migration through the cell membrane. The model considers the interstitial space defined by the transport protein subunits to be the migration pathway for polar solutes. A Langmuir-type adsorption equilibrium is assumed at the interfaces and a multicomponent diffusion mechanism of solute and water is postulated within the migration pathway, where the polar residues of the transport protein represent another component of the system. Membrane selectivity is governed by the adsorption constants, which are shown to affect strongly the kinetics of transport. Isosmotic transport and the volume change of the cell are important features incorporated in the model, which is shown to fulfill the peculiar properties of facilitated diffusion systems. It is concluded that the same type of pathway can be used for the transport of other polar solutes through existing or induced hydrophilic channels, for which a similar approach is suggested.