Temperature-regulated expression of the tac/lacl system for overproduction of a fungal xylanase in Escherichia coli

Temperature-regulated expression of recombinant proteins in the tac promoter (Ptac) system was investigated. Expression levels of fungal xylanase and cellulase from N. patriciarum in E. coli strains containing the natural lacI gene under the control of the Ptac markedly increased with increasing cultivation temperature in the absence of a chemical inducer. The specific activities (units per milligram protein of crude enzyme) of the fungal xylanase and cellulase produced from recombinant E. coli strain pop2136 grown at 42 degrees C were about 4.5 times higher than those of the cells grown at 23 degrees C and were even slightly higher when compared with cells grown in the presence of the inducer isopropyl beta-D-thiogalactopyranoside. The xylanase expression level in the temperature-regulated Ptac system was about 35% of total cellular protein. However, this system can not be applied to E. coli strains containing lacIq, which confers over production of the lac repressor, for high-level expression of recombinant proteins. In comparison with the lambda PL system, the Ptac-based xylanase plasmid in E. coli pop2136 gave a considerably higher specific activity of the xylanase than did the best lambda PL-based construct using the same thermal induction procedure. The high-level expression of the xylanase using the temperature-regulated Ptac system was also obtained in 10-litre fermentation studies using a fed-batch process. These results unambiguously demonstrated that the temperature-modulated Ptac system can be used for overproduction of some non-toxic recombinant proteins.     

Mineral nutrition of slash pine in subtropical Australia. II. foliar nutrient response to fertilization

In the previous paper, we reported the stand growth of slash pine (Pinus elliottii) during the first 11.5 years of plantation in response to (1) initial fertilization at plantation establishment with P rates of 11, 22, 45 and 90 kg P ha^–1 which were either banded or broadcast in the presence or absence of basal fertilizers containing 50 kg N ha^–1, 50 kg K ha^–1 and 5 kg Cu ha^–1 and (2) application of additional 40 kg P ha^–1 at age 10 years. Here we present the responses in foliar nutrient concentrations of slash pine in the first 11.5 years to the initial fertilization and the additional P applied at age 10 years.Foliar N and K concentrations in the first 9.6 years of plantation decreased with the initial P rate. Application of the basal fertilizers improved foliar Cu concentration. Foliar Ca and Mg concentrations increased linearly with the initial P rate. The initial fertilization did not affect foliar Mn concentration in the first 9.6 years. Foliar P concentration increased quadratically with the initial P rate, which accounted for 77–86% of the variation in foliar P concentration. Most of the explained variation in foliar nutrient concentrations was attributable to the plantation age except for foliar P concentration. In the case of foliar P concentration, 53% was explained by the initial P rate, 31% by the plantation age and 2% by the positive interaction between the initial P rate and the plantation age. Foliar P concentration of slash pine at age 11.5 years increased quadratically with the initial P rate and linearly with the additional 40 kg P ha^–1 applied at age 10 years, accounting for 81% of the variation in the foliar P concentration. Foliar nutrient analysis indicated that P was the major limiting nutrient affecting the stand growth of slash pine in the first 11.5 years.     

聚酯酰胺热熔胶的合成及其性能研究

研究了以聚酯胶和聚酰胺胶为原料在催化剂作用下加热共熔,通过大分子链交换反应制备织物用聚酯酰胺热熔胶的方法。用聚酯胶的溶剂对所得产品进行溶取试验并且通过红外光谱分析都证明了链交换反应的存在。用该法所得产品粘接强度大,既耐水洗又耐干洗,适用于粘合衬生产。     

Transformation of fatty acid hydroperoxides by alkali and characterization of products

It has previously been determined that (13S,9Z,11E)-13-hydroperoxy-9,11-octadecadienoic acid was mainly converted into (13S,9Z,11E)-13-hydroxy-9,11-octadecadienoic acid by 5 N KHO with preservation of the stereochemistry of the reactant [Simpson, T.D., and Gardner, H.W. (1993)Lipids 28, 325–330]. In addition, about 20–25% of the reactant was converted into several unknown by-products. In the present work it was confirmed that the stereochemistry was conserved during the hydroperoxy-diene to hydroxydiene transformation, but also, novel by-products were identified. It was found that after only 40 min reaction (9Z)-13-oxo-trans-11,12-epoxy-9-octadecenoic acid accumulated to as much as 7% of the total. Later, (9Z)-13-oxo-trans-11,12-epoxy-9-octadecenoic acid began to disappear, and several other compounds continued to increase in yield. Two of these compounds, 2-butyl-3,5-tetradecadienedioic acid and 2-butyl-4-hydroxy-5-tetradecenedioic acid, were shown to originate from (9Z)-13-oxo-trans-11,12-epoxy-9-octadecenoic acid, and they accumulated up to 2–3% each after 4 to 6 h. Some other lesser products included 11-hydroxy-9,12-heptadecadienoic acid, 3-hydroxy-4-tridecenedioic acid, 13-oxo-9,11-octadecadienoic acid and 12,13-epoxy-11-hydroxy-9-octadecenoic acid. Except for the latter two, most or all of the compounds could have originated from Favorskii rearrangement of the early product, (9Z)-13-oxo-trans-11,12-epoxy-9-octadecenoic acid, through a cyclopropanone intermediate.     

E(6) diffraction catastrophe of the primary rainbow of oblate water drops: observations with white-light and laser illumination

Oblate drops of water illuminated perpendicular to their symmetry axis exhibit catastrophe patterns near the primary-rainbow scattering angle. Previous patterns include the hyperbolic umbilic focal section and separate lips events [see, e.g., H. J. Simpson and P. L. Marston, Appl. Opt. 30, 3468 (1991)]. The present observations concern a much higher-order singularity analyzed by J. F. Nye [Proc. R. Soc. London Ser. A 438, 397 (1992)], the E(6) or symbolic umbilic, in the scattering by levitated drops with monochromatic and collimated white-light illumination. Photographs show the colors produced when the drop is illuminated by white light. The E(6) occurs when the Gaussian curvature of the scattered wave front vanishes in both principal directions, resulting in a high degree of directional focusing. This type of focusing, although only slightly explored, is applicable to the more general problem of scattering from penetrable spheroids.     

Evolution of gypsy-group retrotransposons: phylogenetic analysis of domains included in the pol-polyprotein

Transposons of gypsy group are assigned to LTR-containing retrotransposons present in the genomes of invertebrates, fungi, and plants. In this work, a theoretical analysis of the potential products of ORFs of these retrotranposons was conducted. Alignments were obtained and trees of similarity were constructed for domains of the POL region. On the basis of the obtained data, two hypothetically monophyletic subgroups of transposons were distinguished within the framework of the gypsy group, settling the genomes of taxonomically related organisms (the subgroup of "true" gypsy of insects and the subgroup of gypsy-like transposons of plants and fungi). A number of peculiarities of the topology of these trees hypothetically indicate cases of genetic conversion and recombination of domains accompanying the evolution of this group. The amino acid substitution fixation rate was evaluated on the basis of comparison of sequences of the protein products of ORFs. Estimates of the time of divergence of subgroups of gypsy-group transposons are significantly less than estimates of the times of divergence of their host species. One explanation for this discrepancy might be the hypothesis of settlement by transposons of the genomes of isolated host species.