Origin and direction of in vitro replication of Haemophilus ducreyi and Neisseria gonorrhoeae ampicillin resistance plasmids.

The origin of replication of Haemophilus ducreyi and Neisseria gonorrhoeae ampicillin resistance plasmids was located by cloning BamHI restriction fragments into vector plasmid pAT153 and a derivative plasmid, pAT2. Selection was made for plasmid maintenance in a polA mutant. Direction of replication was determined by in vitro replication of plasmid DNA in the presence of radiolabeled deoxynucleotide.     

The effects of crude oil on the growth of Spartinaalterniflora Loisel. and Spartina cynosuroides (L.) Roth

Greenhouse studies were conducted to examine the effects of crude oil on the growth of Spartinaalterniflora Loisel. and S. cynosuroides (L.) Roth from North Carolina. The way in which crude oil came into contact with the plant tissue and/or substratum was an important factor in determining the responses of both species to oil pollution. Plants recovered from a single application of oil to aerial tissue with relatively little impact on productivity. The presence of an oil layer on the surface of an overlying layer of water had little impact on existing aerial portions of S. alterniflora plants; however, regrowth following harvest was completely inhibited. Incorporation of oil into the substratum significantly reduced aerial productivity and regrowth of S. alternflora and S. cynosuroides. Observations suggest that decreased productivity and regrowth may have been caused by decreased root and rhizome growth. Regrowth potential of S. alterniflora grown in oiled substratum was greater in fine-textured marsh substratum than in sand substratum.     

An isoelectric variant of the 150,000-dalton neurofilament polypeptide. Evidence that phosphorylation state affects its association with the filament

A soluble isoelectric variant of the 150,000-dalton neurofilament protein was isolated from bovine brain by treating a partially purified filament preparation with a low-ionic-strength high-pH buffer. The protein (S150) had similar peptide maps to the neurofilament component of the same molecular weight (NF150) and was recognized by a polyclonal antibody made against the NF150 polypeptide. However, only half the anti-NF150 activity could be removed with the S150 protein. In addition, the S150 protein had a higher isoelectric point than the NF150 protein. Phosphate analysis indicated that the S150 protein was considerably lessened in phosphate content, which could account for the higher isoelectric point of the protein. It appears, therefore, that the S150 protein may be a precursor of NF150 or the result of phosphatase activity during the isolation procedure. Assembly studies showed that the S150 protein, unlike the NF150 protein, could not assemble with the 70-kDa neurofilament protein, indicating that the phosphate groups which were removed are important in the association of this protein to the neurofilament. When filaments containing all three triplet neurofilament polypeptides or those composed of the 70- and 150-kDa neurofilament proteins were subjected to acid phosphatase, a soluble fraction was obtained, which contained isoelectric variants with higher pI values than the NF150 polypeptide. Only unmodified NF150 protein was found in the insoluble fraction. These results support the argument that removal of phosphate groups results in the dissociation of this protein from the filament.     

Replication of adenovirus mini-chromosomes

We have isolated adenovirus origins of DNA replication from both the right and left ends of the genome, which are functional on linear autonomously replicating mini-chromosomes. The mini-chromosomes contain two cloned inverted adenovirus termini and require non-defective adenovirus as a helper. Replicated molecules are covalently attached to protein, and DNA synthesis is initiated at the correct nucleotide even when the origins are not located at molecular ends. The activity of embedded origins leads to the generation of linear minichromosomes from circular or linear molecules. These observations therefore suggest that sequences within the adenovirus origin of replication position the protein priming event at the adenovirus terminus. Experiments investigating the regeneration of deleted viral inverted terminal repeat sequences show a sequence-independent requirement for inverted sequences in this process. This result strongly suggests that repair results from the formation of a panhandle structure by a displaced single strand. On the basis of these observations we propose a model for the generation of adenovirus mini-chromosomes from larger molecules.     

Biotic regionalization and climatic controls in western North America

New methods of weather analysis accompanied by microhabitat ‘bioassays’ have been applied in several case studies to demonstrate effects of atmospheric processes on patterns of community composition and structure and potential species evolution. Average spatial and seasonal airmass dynamics which determine regional and elevational patterns of relative microhabitat favorability, were found to vary between a recent global warming trend (ca 1900 to 1940) and the subsequent global cooling trend (ca 1940 to 1970). These apparently systematic spatial and temporal shifts in weather were related to plant establishment patterns and community composition and structure. The proposed causal mechanisms function, in part, through regional shifts in microhabitat size. These effects are similar to larger scale, longer term shifts deduced from the late Quaternary fossil record. By modifying the spatial approach, month-to-month and year-to-year variability of weather has been examined for the last 130 years at individual points in southwestern North America. Three climatic regimes (the end of the Little Ice Age, the recent warming trend and the recent cooling trend) exhibited distinct year-to-year patterns of weather that can be related to the establishment of different kinds of plants (e.g., C₄ grasses versus C₃ shrubs). Oscillations between different temporal climatic regimes appear to promote the episodic establishment of different life forms, but not necessarily their local extinction. The two methods of weather analysis have been combined in a regional assessment of climatic controls of different biomes in space and time with a primary focus on the Chihuahuan desert. Natural ecotones between the Chihuahuan desert and neighboring biomes are clearly related to large scale airmass dynamics associated with seasonal oscillations in jetstream position. The weather patterns controlling ecotonal positions result from seasonal topographic influences on the general circulation of the atmosphere. The apparent stability of these patterns allows causal hypotheses of biogeographic dynamics and the evolution of physiological traits and life history characteristics.     

High-cell-density fermentation studies of a recombinantE. coli that expresses atrial natriuretic factor

Summary Studies are presented on the fermentation of recombinantEscherichia coli that express rat atrial natriuretic factor (ANF) as a fusion protein. Our objective was to achieve high cell density while maintaining ANF expression at the same level as observed in shake flasks. Improved fermentation conditions included: maintaining glucose concentrations at 1 g/l, using an enriched medium, adding concentrates of medium throughout the fermentation, and blending oxygen for adequate aeration. Cell densities of 12 g/l (dry weight) were achieved, which represented a 10-fold increase over non-improved conditions, while maintaining ANF levels at 7 mg/g of dry cell mass. When galactose was used as an initial carbon source or as a feed supplement, there was a 2-3-fold increase in the expression of ANF from these high-cell-density fermentations. The recombinant ANF was biologically active.     

Polymorphisms within the HLA-DR3 haplotypes

HLA-DR molecules were isolated from eight different HLA-DR3 homozygous B-cell lines by immunoprecipitation with monoclonal antibodies, and they were subsequently analyzed by two-dimensional gel electrophoresis. We found that HLA-DR3 homozygous B-cell lines of consanguineous origin express two types of HLA-DR molecules. One type of HLA-DR molecule was present in all the cell lines tested, whereas the second DR molecule appears to be polymorphic. DNA isolated from the different HLA-DR3 homozygous cell lines was studied by Southern blot analysis to determine whether any DR restriction fragment length polymorphism could be observed. Polymorphisms detected at both the product and genomic level have been compared to each other, and their relations to the serological (HLA-DR) and cellular (HLA-D and LB-Q1) typing data will be discussed.No reprints available     

Microbial-invertebrate interactions and potential for biotechnology

Conclusion As the interactions between marine invertebrates and their bacterial commensals and symbionts are better understood, the application of biotechnology will enhance both environmental and economic benefit. In the immediate future, marine bacteria, either selected or genetically engineered, will play a significant role in enhancing the development of selected invertebrates in aquaculture and in the field. Luck may also favor discovery of mechanisms to suppress the development of biofouling species, perhaps by making it possible to coat submerged surfaces with bacterial films designed to repell larvae and/or interfere with larval morphogenesis. In any case, the future is appealing.     

Temporal relationship between the deposition and microbial degradation of lignocellulosic detritus in a Georgia salt marsh and the Okefenokee Swamp

Temperature dependence and seasonal variations in rates of microbial degradation of the lignin and polysaccharide components of specifically radiolabeled lignocelluloses were determined in sediment and water samples from a Georgia salt marsh and the nearby Okefenokee Swamp. Although temperature regimes in the two ecosystems were similar, rates of mineralization ofSpartina alterniflora lignocellulose in salt marsh sediments increased eightfold between winter and summer, whereas rates of mineralization of lignocellulose from an analogous freshwater macrophyte,Carex walteriana, in Okefenokee sediments increased only twofold between winter and summer. Temperature was the major factor influencing seasonal variations in rates of lignocellulose degradation in both environments. At any given temperature, no substantial differences in lignocellulolytic potential were observed with sediment samples collected at each season. In both ecosystems, the bulk of the lignocellulosic detritus was not degraded at the time of its peak deposition during the fall and winter. Instead, the periods of maximal decomposition occurred during the following spring and summer. These results suggest that periods of maximal nutrient regeneration from the mineralization of lignocellulosic detritus coincide with periods of highest primary production, and that, depending on hydrologic conditions, significant horizontal transport of essentially intact lignocellulosic material is possible due to the lag period between deposition and microbial degradation.