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Release of a chimeric protein into the medium from Escherichia coli using the C-terminal secretion signal of haemolysin. 总被引:29,自引:6,他引:23
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Recently, we have identified a novel topogenic sequence at the C terminus of Escherichia coli haemolysin (HlyA) which is essential for its efficient secretion into the medium. This discovery has introduced the possibility of using this secretion system for the release of chimeric proteins from E. coli directly into the medium. We have now successfully fused this C-terminal signal to a hybrid protein containing a few residues of beta-galactosidase and the majority of the E. coli outer membrane porin OmpF lacking its own N-terminal signal sequence. We find that this chimeric protein is specifically translocated across the inner and outer membranes and is released into the medium. In addition, we have further localized the HlyA secretion signal to the final 113 amino acids of the C terminus. In fact, a specific secretion signal appears to reside at least in part within the last 27 amino acids of HlyA. 相似文献
4.
Joseph C. Polacco Adam K. Judd Jody K. Dybing Silvia R. Cianzio 《Molecular & general genetics : MGG》1989,217(2-3):257-262
Summary We reported earlier the recovery of two classes of soybean urease mutants in soybean (Glycine max L. Merr. cv. Williams). Class I mutants lack the embryo-specific urease while class II mutants lack the activities of both
urease isozymes, the embryo-specific and the ubiquitous urease, the latter found in all tissues examined. We report here the
recovery of a true-breeding mutant, aj3, which represents the third phenotypic class: normal levels of embryo-specific urease
and little or no ubiquitous urease. Unlike class II mutant plants which lack urease in all tissue, aj3 lacks urease activity
only in leaves (ca. 2% normal activity); its roots have near normal urease activity. Callus derived from leaves of aj3 has
14% to 40% the urease activity of Williams 82 callus. This partial reduction in urease activity in aj3 callus is sufficient
to reduce growth with urea as sole nitrogen source and to confer resistance to 50 mM urea added to callus maintenance medium.
Leaves of aj3 produce more than 40 times the urease antigen expected from their urease activity. The aj3 trait is due to a
single recessive lesion which is not allelic with lesions at theEu2, Eu3 (class II) orEu1 (class I) loci. We designate the aj3 genotype aseu4/eu4. 相似文献
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The isolation and immunolocalization of iron-binding compounds produced by Gloeophyllum trabeum 总被引:2,自引:0,他引:2
Jody Jellison Vikas Chandhoke Barry Goodell Frank A. Fekete 《Applied microbiology and biotechnology》1991,35(6):805-809
Summary Low molecular weight iron-binding compounds are produced by the brown-rot fungus Gloeophyllum trabeum. These chelators may function in scavenging transition metals for fungal metabolism and extracellular enzyme production. Because of the low molecular mass of the chelate-metal complex (below 1000 Da), and the oxidizing potential of the bound transition metals, certain chelating compounds could also play a role in the early stages of cellulose depolymerization by brown-rot fungi. High-affinity iron-binding compounds were isolated and partially purified from both liquid cultures of the brown-rot Gloeophyllum trabeum and from infected wood. Chelating compounds purified by thin-layer chromatography were used to prepare specific antibodies. These antibodies were shown to detect the chelator in infected wood and liquid fungal cultures by enzyme-linked immunosorbent assay and could be used in immunotransmission electron microscopy to visualize the high-affinity iron-binding compounds in situ. Elucidating the physiological roles of fungal chelate-metal complexes and determining their function in lignocellulose depolymerization will help us to better understand the mechanism of wood biodegradation.Publication no. 1549 Maine Agricultural Experiment Station
Offprint requests to: J. Jellison 相似文献
7.
Female white-footed mice (Peromyscus leucopus) fail to produce offspring when paired with a male from weaning until 150 days of age if an adult female or her odor is also present. The present study delineates more clearly which stage of the young female's reproductive cycle is inhibited by the chemosignal of the older female. Age at vaginal opening and first estrus are delayed by the presence of the older female, but only for about 10 days. The presence of a male counteracts this delay. Thus, the basis for the block is not a delay in puberty. The older female's presence does not influence the number of estrous cycles experienced during the 30 days following first estrus, nor does it influence the number of corpora lutea found at autopsy. The presence of sperm in the vaginae of young females indicates that they were copulating. Likewise, examination of embryos 2 and 3 days after copulation reveals normal developmental progress. However, implantation does not occur in young females that have been exposed to an adult female. Thus, the block occurs either during the final stages of embryo transport or in relation to the implantation process itself. 相似文献
8.
Observations of Barophilic Microbial Activity in Samples of Sediment and Intercepted Particulates from the Demerara Abyssal Plain 总被引:4,自引:4,他引:0
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To better understand the ecological significance of pressure effects on bacteria in the abyssobenthic boundary layer, experimental suspensions of sediments and sinking particulates were prepared from samples collected in boxcore and bottom-moored sediment traps at two stations (depth, 4,470 and 4,850m) in the Demerara abyssal plain off the coast of Brazil. Replicate samples were incubated shipboard at 3°C and at both atmospheric and deep-sea pressures (440 or 480 atm [4.46 × 104 or 4.86 × 104 kPa]) following the addition of [14C]glutamic acid (<10 μg liter−1) or yeast extract (0.025%) and the antibiotic nalidixic acid (0.002%). In seven of the eight samples supplemented with isotope, a barophilic microbial response was detected, i.e., substrate incorporation and respiration were greater under in situ pressure than at 1 atm (101.3 kPa). In the remaining sample, prepared from a sediment trap warmed to 24°C before recovery, pressure was observed to inhibit substrate utilization. Total bacterial counts by epifluorescence microscopy decreased with depth in each sediment core, as did utilization of glutamic acid. Significant percentages of the total bacterial populations in cold sediment trap samples (but not the prewarmed one or any boxcore sample) were abnormally enlarged and orange fluorescing after incubation with yeast extract and nalidixic acid under deep-sea conditions. Results indicated that in the deep sea, barophilic bacteria play a predominant role in the turnover of naturally low levels of glutamic acid, and the potential for intense microbial activity upon nutrient enrichment is more likely to occur in association with recently settled particulates, especially fecal pellets, than in buried sediments. 相似文献
9.
Targeted Selection Experiments and Enzyme Polymorphism: Negative Evidence for Octanoate Selection at the G6PD Locus in DROSOPHILA MELANOGASTER 总被引:4,自引:3,他引:1
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Published studies have reported significant selection with respect to the G6pd locus for Drosophila melanogaster reared on Na-octanoate food. We have reexamined the selective effects of Na-octanoate on egg to adult viability with respect to the G6pd polymorphism using specially constructed X chromosomes. Four experiments were carried out using different 6Pgd backgrounds in two recombinant sets of chromosomes segregating for the G6pd locus but constructed so as to minimize variation over most of the X chromosome. In addition, two measures of viability were used, and the size of the experiments and their associated degrees of freedom are approximately double those reported in the former studies. Our results find no evidence for differential selection on G6pd genotypes (males and females) by Na-octanoate and, therefore, do not corroborate the positive results of selection reported by other investigators. The reasons for our different results are discussed. 相似文献
10.
The hitch-hiking effect of a favourable gene 总被引:70,自引:0,他引:70