Gene fusion vectors based on the gene for staphylococcal protein A

Two plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusion, have been constructed. These vectors will allow fusion of any gene to the protein A gene, thus giving hybrid proteins which can be purified, in a one-step procedure, by IgG affinity chromatography. As an example of the practical use of such vectors, the protein A gene has been fused to the lacZ gene of Escherichia coli. E. coli strains containing such plasmids produce hybrid proteins with both IgG binding and β-galactosidase activities. The hybrid protein(s) can be immobilized on IgG-Sepharose by its protein A moiety with high efficiency without losing its enzymatic activity and they can be eluted from the column by competitive elution with pure protein A. The fused protein(s) also binds to IgG-coated microtiter wells which means that the in vivo product can be used as an enzyme conjugate in ELISA tests.     

Effects of electroconductive heat treatment and electrical pretreatment on thermal death kinetics of selected microorganisms

Suspensions of yeast cell (zygo Saccharomyces bailii) in a phosphate buffer solution were subjected to conventional (hot water) and ohmic (electric current) heating under identical temperature histories. Experiments were also conducted with cells of Escherichia coli to compare the lethal effect of combination of sublethal electrical preteatment and conventional heating with conventional heating. The kinetic parameters (D,Z,K and E(a)) were determined for both organisms during different treatments. There was no significant difference in the death rate of yeast cells during conventional and ohmic heating at the voltage range used in this study. Results of electrical pretreatment and conventional heating on E. coli indicated differences under certain conditions when compared with pure conventional heating. Thus it is concluded that microbial death during ohmic heating was due primarily to thermal effects with no significant effect of electric current per se. Sublethal electrical pretreatment appears to offer potential for increased bacterial inactivation in certain cases.     

Probing nucleosome core secondary structure before and after α-chymotrypsin treatment by raman spectroscopy and thermal denaturation

Nucleosome cores were digested with α-chymotrypsin until histone H3 was degraded to a partial histone, CP1. As we reported previously, cleavage occurred at leucine 20 to H3 and resulted in an increase in circular dichroism between 265 to 285 nm. Some modest core unfolding was also observed as determined by a small decrease in the sedimentation coefficient. Studies reported here deal with the analysis of core secondary structure and subsequent perturbation caused by treatment with α-chymotrypsin. Raman spectroscopy indicated that chymotryptic treatment promoted a change in the conformational environment of a population of core histone tyrosines. In addition, a shift from B-form to an intermediate B- or A-form was observed for core DNA. High-resolution thermal denaturation was used to determine alterations in the stabilization of core DNA related to perturbation of the core histones. Brief chymotryptic treatment indicated changes in both pre-melt and irreversible transitions.     

Thermal acclimation and hardening in tadpoles of the bullfrog, Rana catesbeiana

1. 1.|Critical thermal maxima (CTMax) and minima (CTMin) were measured to evalute thermal hardening in Rana catesbeiana.

2. 2.|Tadpoles show heat hardening and CTMax acclimation, and both responses are influenced by developmental stage.

3. 3.|The first evidence of cold hardening in vertebrates is reported here.

4. 4.|Heat hardening significantly reduces cold tolerance, but there is otherwise no evidence of a cross-hardening effect.

Rate of acclimation of the tropical salt-marsh fish Cyprinodon dearborni to temperature changes

The acclimation rates of temperature changes in Cyprinodon dearborni, collected from Laguna Los Patos, Cumana, Venezuela, were determined by the critical thermal maximum method. At an increase in temperature (from 24 to 31°C) fish started to gain acclimation level after 3 hours and took 3 days to fully get up to a higher level of resistance to heat death; however, at a decrease in temperature (from 3 t to 24°C) fish began to lose its acclimation level after 12 days and required 39 days to reach a lower level of resistance to thermal death.     

The rotifer fauna of the River Loire (France), at the level of the nuclear power plants

In the heated affluents of nuclear power plants of the river Loire, rotifers are abundant. Cosmopolitan species are numerically dominant, but a tropical fauna is also present, among which the genus Brachionus, representing 20% of the total species, is best represented. From a comparison between water upstream and downstream of the power plants, it further appears that in downstream warmed-up waters, some species show an important development, but not in colder upstream waters.     

The influence of coal ash and thermal discharges upon the distribution and bioaccumulation of aquatic invertebrates

The distribution, density and uptake of twenty elements by aquatic invertebrates inhabiting a drainage system, that received excessive coal ash effluent (275 JTU of turbidity) at one end and thermal loading (44.5°C) at the other end, was studied for 15 months. The ash settling basin filled during the first eight months of sampling which resulted in the release of ash effluent directly into the receiving system. Density of invertebrates was lowest in the 300 m stream between the ash basin and swamp and highest 1200 m beyond the stream-swamp confluence where ash influence was minimal. Invertebrate density was lowest in the stations where turbidity from ash effluent was greatest. The most tolerant invertebrates to coal ash stress were odonates (Libellula sp. and Enallagma sp.), crayfish (Procambarus sp.), amphipods (Gammarus sp.) and gastropods (Physa. sp.), and midges (Chironomidae) when the basin was filling. During the period of ash overflow, all groups were either reduced in numbers or absent. In the thermally stressed station, Libellula sp. was the predominant invertebrate sampled when water temperature ranged from 25.5–45.5°C (257-1=28.7°C) all aquatic invertebrates were limited in numbers and density when temperature exceeded the lower and upper ranges of 10.0–38.0°C.This research was supported by AEC Contract AT (38-1-824)This research was supported by AEC Contract AT (38-1-824)