Suppression by the ColV,I-K94 plasmid of inhibitor sensitivities in ompA mutants of Escherichia coli

A series of ompA mutants derived from Escherichia coli K12 strains showed increased sensitivity (compared with the ompA+ parents) to aminoglycoside antibiotics and to other cationic agents including polymyxin B. One tested mutant also showed increased sensitivity to nafcillin and fusidic acid, but not to the hydrophilic ampicillin. All these inhibitor sensitivities in the ompA mutants were suppressed by ColV, I-K94 and by certain other ColV plasmids, but not by any of the other tested large plasmids. Suppression correlated with the production of the VmpA protein, but transfer and colicin components were not needed for suppression. Further comparison of the ompA and vmpA genes and their products was made and it indicated that there is little if any homology between the genes, that the synthesis of their products is regulated by quite different mechanisms, and that regions of these gene products exposed at the cell surface show different susceptibility to protease attack after denaturation.     

Evidence that the phytochrome gene family in black cottonwood has one PHYA locus and two PHYB loci but lacks members of the PHYC/F and PHYE subfamilies

The phytochrome photoreceptors play important roles in the photoperiodic control of vegetative bud set, growth cessation, dormancy induction, and cold-hardiness in trees. Interestingly, ecotypic differences in photoperiodic responses are observed in many temperate- zone tree species. Northern and southern ecotypes of black cottonwood (Populus trichocarpa Torr. & Gray), for example, exhibit marked differences in the timing of short-day-induced bud set and growth cessation, and these responses are controlled by phytochrome. Therefore, as a first step toward determining the molecular genetic basis of photoperiodic ecotypes in trees, we characterized the phytochrome gene (PHY) family in black cottonwood. We recovered fragments of one PHYA and two PHYB using PCR-based cloning and by screening a genomic library. Results from Southern analyses confirmed that black cottonwood has one PHYA locus and two PHYB loci, which we arbitrarily designated PHYB1 and PHYB2. Phylogenetic analyses which included PHY from black cottonwood, Arabidopsis thaliana and tomato (Solanum lycopersicum) suggest that the PHYB/D duplications in these species occurred independently. When Southern blots were probed with PHYC, PHYE, and PHYE heterologous probes, the strongest bands that we detected were those of black cottonwood PHYA and/or PHYB. These results suggest that black cottonwood lacks members of the PHYC/F and PHYE subfamilies. Although black cottonwood could contain additional PHY that are distantly related to known angiosperm PHY, our results imply that the PHY family of black cottonwood is less complex than that of other well-characterized dicot species such as Arabidopsis and tomato. Based on Southern analyses of five black cottonwood genotypes representing three photoperiodic ecotypes, substantial polymorphism was detected for at least one of the PHYB loci but not for the PHYA locus. The novel character of the PHY family in black cottonwood, as well as the differences in polymorphism we observed between the PHYA and PHYB subfamilies, indicates that a number of fundamental macro- and microevolutionary questions remain to be answered about the PHY family in dicots.      

Decreased DNA damage by acid and increased repair of acid-damaged DNA in acid-habituated Escherichia coli

A study of the conjugal transfer of ColV,I-K94 tn 10 from acid-treated donors suggested that acid-habituated recipients repair acid-damaged plasmid DNA better than those that are not habituated. The presence of an increased repair activity for acid-damaged DNA in habituated cells was confirmed by isolating pBR322 from acid-treated organisms; habituated cells produced more transformants when transformed by it than did non-habituated ones. Additionally, agarose gel electrophoretic studies of pBR322 DNA isolated from acid-damaged cells and tests of its transforming activity both indicated that plasmid DNA in habituated cells is less damaged by extreme acidity than is that in non-habituated organisms.     

Molecular cloning and expression of human Ca(2+)-sensitive cytosolic phospholipase A2

Phospholipases A2 (PLA2s) play a key role in inflammatory processes through production of precursors of eicosanoids and platelet-activating factor. Recently, we described the purification of a novel approximately 100-kDa cytosolic PLA2 (cPLA2) from human monoblast U937 cells that is activated by physiological (intracellular) concentrations of Ca2+ (Kramer, R. M., Roberts, E. F., Manetta, J., and Putnam, J. E. (1991) J. Biol. Chem. 266, 5268-5272). Here we report the isolation of the complementary DNA encoding human cPLA2 and confirm its identity by expression in bacteria and in hamster cells. The predicted 749-amino acid cPLA2 protein has no similarity to the well known secretory PLA2s, but contains a structural element homologous to the C2 region of protein kinase C. The molecular cloning of cPLA2 will allow further studies defining the structure, function, and regulation of this novel PLA2.     

Regulation of purified rat liver acetyl CoA carboxylase by phosphorylation

Acetyl CoA carboxylase was purified from liver of fasted-refed rats to near homogeneity, based on electrophoretic analysis and biotin content. These preparations contained an endogenous protein kinase that catalyzed the transfer of radioactive phosphate from [gamma-32P]ATP to acetyl CoA carboxylase, accompanied by a decrease in acetyl CoA carboxylase activity. Phosphate incorporated into acetyl CoA carboxylase was removed when the preparation was incubated with partially purified phosphorylase phosphatase catalytic subunit with regain of enzymatic activity. This endogenous protein kinase was shown not to be affected by either cyclic-AMP-dependent protein kinase inhibitor, EGTA, or trifluoperazine. The addition of either cyclic-AMP or purified cyclic-AMP-dependent protein kinase catalytic subunit to the purified acetyl CoA carboxylase preparation increased protein phosphorylation but had no further effect on acetyl CoA carboxylase activity. Purified acetyl CoA carboxylase was shown to act as an ATPase during the phosphorylation reaction.     

牛脾转移因子的提取工艺研究

目的：比较从牛脾中提取活性ＴＦ的不同方法。方法：分别利用Ｌａｗｒｅｎｃｅ－－透析法、Ｌａｗｒｅｎｃｅ－超滤法、超离心－超滤法提取,再以蛋白反应、红外及紫外扫描予以鉴定比较。结果：Ｌａｗｒｅｎｃｅ－超滤法和超离心－超滤法效果较好。结论：工业化生产中应采用Ｌａｗｒｅｎｃｅ－超滤法。     

Extracellular proteins and other components as obligate intermediates in the induction of a range of acid tolerance and sensitisation responses in Escherichia coli

Several acid tolerance responses of Escherichia coli were associated with secretion into the growth media of components (frequently proteins) which altered acid tolerance of other cultures. First, medium filtrates from cultures induced to acid tolerance by several conditions converted pH 7.0-grown organisms to tolerance and, for most such responses, filtrate proteins were needed for full induction. Secondly, filtrates from cultures induced to acid sensitivity at alkaline pH produced sensitisation of resistant cultures. Thirdly, filtrates from inherently tolerant or sensitive strains altered tolerance or sensitivity of normal strains. In many cases, filtrate components were essential for the original response e.g. acid habituation at pH 5.O. Extracellular components may function as intermediates only in stress tolerance responses, but other adaptive responses must be tested as such components may function in other inducible processes.     

Symbiotic dinoflagellates in marine Cnidaria: diversity and function

Dinoflagellates of the genus Symbiodinium are the most common symbiotic algae in benthic marine Cnidaria. This review addresses our current understanding of the molecular diversity of Symbiodinium and the function of these algae in symbiosis. Ribosomal DNA sequence data indicate that Symbiodinium is a diverse but probably monophyletic group. They also provide a phylogenetic framework for the analysis of the functional diversity of Symbiodinium (i.e. the variation in phenotype among various Symbiodinium genotypes), especially in relation to their nutritional role in the symbiosis. Symbiodinium provides the animal host with photosynthetic carbon and may also recycle animal nitrogenous waste. These interactions are advantageous to animals in shallow, oligotrophic waters. Recent developments in understanding of both photosynthate release and nitrogen relations in the symbiosis are reviewed. They provide the basis to explore the variation in nutritional interactions among different Symbiodinium genotypes. This review highlights areas of current uncertainty and controversy and addressess possible fulture directions of research.     

Effects of an abrupt diet change from hay to concentrate on microbial numbers and physical environment in the cecum of the pony

Microbial numbers, pH, fluid volume, and turnover rate in the pony cecum were measured during an abrupt change from an all-forage to an all-concentrate diet, both fed at maintenance energy levels. Concentrate feeding resulted in increased (P less than 0.01) numbers of total viable anaerobic bacteria. The numbers of organisms growing on selective starch medium increased (P less than 0.01) when concentrate was fed, while numbers on xylan and pectin media decreased (P less than 0.025). Seven days after the diet change to concentrate, the number of bacteria growing on lactate medium increased (P less than 0.01), followed by a gradual decline. Cellulolytic bacteria occurred in low numbers, ranging from 1.1 x 10(4) to 4.4 x 10(4) per g of cecal contents. Feeding all concentrate decreased both the number of genera (P less than 0.01) and total protozoan numbers (P less than 0.01) in the cecum. Minimum cecal pH values of 6.4 and 5.8 were obtained when forage and concentrate, respectively, were fed, with the minimum pH occurring 6 h postfeeding. Dry-matter percentage of cecal contents followed a diurnal pattern which was the inverse of the pH curve. During forage feeding, the cecum contained an average of 2.2 liters (1.6 to 3.4 liters), which turned over 3.9 times per day. When concentrate was fed, cecal volume averaged 3.9 liters (0.6 to 8.6 liters), with a mean liquid turnover of 4.2 times per day. Microbial numbers and pH changes in the pony cecum associated with an abrupt change in diet from hay to concentrate resembled those which occur in the rumen under similar feeding conditions.