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1.
Nutritional Factors Influencing the Development of Competence in the Bacillus subtilis Transformation System 总被引:36,自引:31,他引:5
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Cultures of Bacillus subtilis developed competence for the uptake of deoxyribonucleic acid in a chemically defined medium with a predictable, reproducible pattern. The gross effects of individual amino acids were determined. Seven amino acids, most of which are reported to be major components of the cell wall, were shown to impair the development of maximal levels of competence. When the synthetic growth medium was supplemented with a mixture of the nine amino acids which we found to stimulate the development of competence, the level of transfection was increased to 10 to 15% of the population. The actual level of competence in these populations was assayed by transformation of unlinked bacterial markers and by two different transfection assays. The results indicate that calculations from cotransfer of unlinked markers overestimates the degree of competence in highly competent populations of B. subtilis, whereas the number of plaques obtained in transfection is an under-estimate of the actual level of competence. The results are interpreted to indicate that neither method of analysis gives a true estimate of the competent population, but that more than 80% of the cells may be competent. 相似文献
2.
The Role of Competence in the Cranio-Caudal Segregation of the Central Nervous System 总被引:5,自引:5,他引:0
Left to right thirds of Triturus presumptive prosencephalon show identical developmental potencies after implantation in a neutral Ambystoma environment. Such equipotential grafts were excised from stages between late-gastrula and mid-neurula and implanted into the neural plate of an Ambystoma host at different cranio-caudal levels. Their regional differentiation was independent of the age of the host, but dependent upon the age of the donor material; the older the latter the smaller the portion of the graft which was transformed into more posterior neural structures. Full transformation occurred in stage 11/12 grafts, while pure prosencephalic differentiation took place in stage 16 grafts, demonstrating that the period of competence of the neurectoderm for transformation extends from stage 11/12 up to stage 16. Irrespective of the level of implantation all grafts older than stage 11/12 and younger than stage 16 showed an uninterrupted cranially-oriented regional differentiation. The medio-lateral extension of the transformation process is primarily determined by the temporal loss of competence of the implanted neurectoderm. A comparison of grafts implanted at different cranio-caudal levels showed that transformation is more pronounced the more caudal the level of implantation, so that another factor(s) than competence must also play a role in the regional segregation of the CNS. 相似文献
3.
Summary Evidence of early loss of immunological competence in cases of neoplasms occurring in juveniles was found in an analysis of OSCC data (Oxford Survey of Childhood Cancers). The effects observed included heightened sensitivity to infection from birth onwards for all types of childhood cancer, higher levels of sensitivity for leukaemia than for lymphomas, and higher levels for lymphomas than for other solid tumours. The findings as a whole are consistent with in utero loss of immunological competence, which is an essential promoter of cancers of foetal origin and thus allows the outcome of an in utero cancer induction to be influenced both by maternal levels of immunological competence and postnatal infection. 相似文献
4.
Development of Competence in the Bacillus subtilis Transformation System 总被引:51,自引:35,他引:16
Competence in Bacillus subtilis, assayed by the ability of cells to be transformed with bacterial deoxyribonucleic acid (DNA) or transfected by phage DNA, has been shown to occur in a single semisynthetic medium with peak activity occurring 3 hr after the cessation of logarithmic growth. No step-down conditions or culture manipulations were necessary for routine transfection of 1% of the population. The results demonstrate that bacteriophage DNA is a valid assay for studying the development of competence in B. subtilis. Predictions of workers using transforming bacterial DNA, who have suggested that competence in B. subtilis is associated with a specific phase of growth, are substantiated. The peak of competence is not affected by marked differences in the rate of growth during the logarithmic phase. The effect on development of competence by this procedure of some components (including casein hydrolysate, tryptophan, and histidine) which were routinely included in the transformation medium by other investigators has been determined by use of infectious phage DNA as an assay. We have demonstrated that tryptophan, as well as histidine, increases the transformation frequency—even in strains which do not have auxotrophic demands for these components. Glutamic acid and alanine depress optimal levels of transfection. 相似文献
5.
Deoxyribonucleic acid (DNA) extracted from phage SPP1 is highly infectious on Bacillus subtilis competent cells; the efficiency of infection is 5 x 10(3) to 6 x 10(3) phage equivalents per plaque-forming unit. This DNA was used to study the relationship between competence for transfection and for transformation. The experiments were concerned with the frequency of infection and transformation in mutants exhibiting different levels of competence, the effect of periodate on competence for infection and for transformation, the competition between phage and bacterial DNA, the transformation of cells preinfected with phage DNA, and the infection of cells pretreated with bacterial DNA. The data show that B. subtilis cells competent for transformation are also competent for transfection and vice versa; transfection with phage DNA represents, therefore, a simple way to measure the total number of competent cells in a culture. The fraction of competent cells, determined by SPP1 DNA infection, varied from 10(-2) to 7 x 10(-2). 相似文献
6.
Full-grown oocytes of Atlantic croaker are insensitive to maturation-inducing steroid (MIS) unless they are primed with gonadotropin (GtH). The objective of this study was to examine the mechanism of GtH-induced maturational competence in croaker oocytes. Specifically, we determined the in vitro secretion of steroids by intact ovarian follicles of unprimed or hCG-primed fish, the direct effects of steroids on maturational competence, and the effects of steroid (cyanoketone), protein (cycloheximide), and RNA (actinomycin D) synthesis inhibitors on hCG-induced maturational competence and steroidogenesis in vitro. The steroid content of the incubation medium after hCG treatment was measured by RIA. The effects of hCG or exogenous steroid treatment on maturational competence were determined by recording the incidence of germinal vesicle breakdown (GVBD) after MIS-induced GVBD in a standard bioassay. Our major findings were: (1) induction of maturational competence occurred after exposure of ovarian follicles to hCG either in vivo or in vitro; (2) MIS secretion was detected in follicles of hCG-primed fish but not unprimed fish, and no MIS secretion was observed during hCG induction of maturational competence in vitro; (3) treatment with cyanoketone blocked the hCG-dependent secretion of testosterone and estradiol but not the development of maturational competence; (4) treatment with MIS or various other exogenous steroids in the absence of hCG did not induce maturational competence; and (5) hCG-induced maturational competence was inhibited by cycloheximide and actinomycin D. Therefore, the mechanisms of GtH induction of oocyte maturation in Atlantic croaker can be described in two distinct stages: a delta-4 steroid-(including MIS) and estrogen-independent priming stage followed by a MIS-mediated GVBD stage. The priming stage may involve mechanisms requiring RNA as well as protein synthesis. 相似文献
7.
ClpP of Bacillus subtilis is required for competence development, motility, degradative enzyme synthesis, growth at high temperature and sporulation 总被引:13,自引:11,他引:2
Tarek Msadek Véronique Dartois Frank Kunst Marie-Laure Herbaud François Denizot & Georges Rapoport 《Molecular microbiology》1998,27(5):899-914
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Spencer, Hugh T. (The Johns Hopkins University School of Hygiene and Public Health, Baltimore, Md.), and Roger M. Herriott. Development of competence of Haemophilus influenzae. J. Bacteriol. 90:911-920. 1965.-A chemically defined nongrowth medium was developed for the induction of competence of Haemophilus influenzae by a stepdown procedure. Cells grown logarithmically in Heart Infusion Broth became competent after being transferred to a medium which consisted of amino acids, sodium fumarate, and inorganic salts. Chloramphenicol (2 mug/ml) or l-valine (1 mug/ml) in the nongrowth medium inhibited development of competence. The inhibitory action of l-valine was reversed by comparable concentrations of l-isoleucine. Kinetic studies of the development of competence showed a variable capacity of competent cells to take up deoxyribonucleic acid and reaffirmed earlier findings that competence was not transmissible in H. influenzae. Addition of nicotinamide adenine dinucleotide, thiamine, calcium pantothenate, uracil, and hypoxanthine to the medium for competence resulted in a minimal growth medium in which reduced levels of competence were developed. 相似文献
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11.
Effect of Competence Induction on Macromolecular Synthesis in a Group H Streptococcus 总被引:6,自引:2,他引:4
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The effects of competence induction by competence factor (CF) on macromolecular synthesis in group H streptococcus strain Wicky were investigated. CF preparations (culture filtrates from competent group H streptococcus strain Challis) were either heated or partially purified to remove a bacteriocin. These preparations did not inhibit growth, although they induced high levels of competence in strain Wicky. The action of the CF preparations did not affect the overall rates of deoxyribonucleic acid and protein synthesis, but caused a reduction in the rates of ribonucleic acid (RNA) and peptidoglycan synthesis. When competence induction by CF was prevented, no alterations in RNA or peptidoglycan synthesis were observed, indicating that these changes are in fact related to the development of competence. 相似文献
12.
The ways of transdifferentiation are considered: spontaneous and induced. Spontaneous transdifferentiation taking place after the disaggregation of cells in the clonal and cell cultures is determined by the competence of the transforming cells themselves. Induced transdifferentiation is determined not only by the competence but also by the effect of external inducing factors. It is suggested that the direction of induced transdifferentiation depends on the ratio between the external and internal inducing factors and on the character of cell cycles. It is probable that the inducing factors entering the cells during the early embryonic induction are reproduced in the cells in a dormant state and some of them do not reveal their presence until appropriate conditions are set. When the cells are isolated in the cultures, the ratio of these factors inside the cell changes and a competence to transdifferentiation is revealed which arises as early as during induction. 相似文献
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Control of bacteriochlorophyll accumulation by light in Rhodobacter capsulatus. 总被引:4,自引:4,他引:0
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A J Biel 《Journal of bacteriology》1986,168(2):655-659
Oxygen levels which control induction of the assembly of the pigment-protein photosynthetic polypeptides in dark-grown Chloroflexus aurantiacus were determined. The induction signal by low-oxygen tension is not directly related to the respiratory competence of these photosynthetic cells. Cytochrome c554, the primary electron donor to P865+ of the reaction center, is not present in dark-grown respiratory cells but is induced in parallel with bacteriochlorophylls a and c and at similar oxygen partial pressure. The development of these components of the photosynthetic apparatus and its electron transport chain is completely independent of the presence of any detectable light or bacteriochlorophyll c or a pigments in C. aurantiacus. 相似文献
16.
Quanli An Wei Peng Yuyao Cheng Zhenzhen Lu Chuan Zhou Yong Zhang Jianmin Su 《Journal of cellular physiology》2019,234(10):17370-17381
Oocyte quality, which is directly related to reprogramming competence, is a major important limiting factor in animal cloning efficiency. Compared with oocytes matured in vivo, in vitro matured oocytes exhibit lower oocyte quality and reprogramming competence primarily because of their higher levels of reactive oxygen species. In this study, we investigate whether supplementing the oocyte maturation medium with melatonin, a free radical scavenger, could improve oocyte quality and reprogramming competence. We found that 10−9 M melatonin effectively alleviated oxidative stress, markedly decreased early apoptosis levels, recovered the integrity of mitochondria, ameliorated the spindle assembly and chromosome alignment in oocytes, and significantly promoted subsequent cloned embryo development in vitro. We also analyzed the effects of melatonin on epigenetic modifications in bovine oocytes. Melatonin increased the global H3K9 acetylation levels, reduced the H3K9 methylation levels, and minimally affected DNA methylation and hydroxymethylation. Genome-wide expression analysis of genes in melatonin-treated and nontreated oocytes was also conducted by high-throughput RNA sequencing. Our results indicated that melatonin ameliorates oocyte oxidative stress and improves subsequent in vitro development of bovine cloned embryos. 相似文献
17.
The endogenous levels of polyamines (PAs) in leaf-base explants isolated from plants of two isogenic lines of Dactylis glomerata L., differing in their competence for somatic embryogenesis, were compared. Leaf-bases isolated from plants with a high level of competence for somatic embryogenesis (HEC) contained four times the level of polyamines compared to those isolated from plants with a low level of competence for somatic embryogenesis (LEC). When the levels of individual polyamines in the HEC and LEC lines were compared, leaf-bases from plants of the HEC line had much lower PUT/SPD ratios than those from the LEC line. When changes in the levels of PAs were monitored during the first 28 d of culture, on a medium which promotes initiation of somatic embryogenesis, leaf-base cultures from plants of the HEC line showed a 50% increase in the levels of PAs during the first 7 d of culture, after which time levels began to decline. By day 21, levels had dropped below those found in freshly isolated leaf bases. While PUT and SPM levels increased by about 30%, the greatest increase was shown by SPD, which increased by more than 100% during the first 7 d of culture, before declining. In contrast much smaller changes in PA levels were found when leaf-bases from plants of the LEC line were cultured. 相似文献
18.
C. Gomez Leonard 《Journal of bacteriology》1973,114(3):1198-1205
Appropriately timed use of trypsin, which inactivates competence factor (CF), and chloramphenicol made feasible a separation and characterization of early events in the development of competence in group H streptococci. Step 1 is production of CF, which is inseparable in time from the concomitant release of free CF into the medium. The producing cells, which are noncompetent at the time, also accumulate cell-bound CF (CB-CF) from the onset of CF synthesis. In step 2, the released CF is adsorbed or taken up in a trypsin-insensitive state by the producing cells and is not destroyed as previously suggested. This occurs rapidly in a transformation-supporting (complete) medium. The rapid decline in free CF is concomitant with the rise in CB-CF, and a maximal increase in the latter does not occur in cultures exposed to trypsin, which inactivates any trypsin-accessible CF. The rapid increase in CB-CF (above trypsin-treated levels) leads to step 3, the induction of competence. All of these steps probably require protein synthesis, because each is inhibited by chloramphenicol. The data also indicate that only free CF that is subsequently adsorbed, and which thus leads to maximal levels of trypsin-insensitive CB-CF, is the effective inducer of competence in either CF-producing (Challis) or CF-nonproducing (Wicky) cultures. The processes induced by the newly bound CF are not fully understood, but certain new properties, previously described by others as indicating competence, were measured during the several steps of competence development. Cell aggregation at pH 2 appears to be related to CB-CF and can be shown before this bound CF has induced competence. The ability of cultures to autolyze maximally can be diminished by trypsin treatment of precompetent cells without affecting subsequent competence development as measured by transformation. 相似文献
19.
Bonser RH Deaton KE Bishop CM Butler PJ 《Journal of experimental zoology. Part A, Comparative experimental biology》2004,301(8):636-641
Thyroid hormones show fluctuating levels during the post-hatching development of birds. In this paper we report the results of the first mechanical tests to quantify the effect of hypothyroidism, during post-natal development, on the skeletal properties of a precocial bird, the barnacle goose, as determined by microhardness testing. The effect of hypothyroidism is tissue-specific; bone from the femora of birds is not significantly affected by induced hypothyroidism, however, there is a strong positive relationship between the levels of circulating thyroid hormones and the mechanical properties of bone from humeri. In the barnacle goose the development of the wing skeleton and musculature depends on an increase in circulating thyroid hormones and our analysis shows that, in its absence, the mechanical competence of the bone mineral itself is reduced in addition to the decreased bone length and muscle development previously reported in the literature. 相似文献
20.
Pamella Akoth Ogada Thomas Debener Hans‐Michael Poehling 《Ecology and evolution》2016,6(21):7911-7920
The complexity of tospovirus–vector–host plant interaction is linked to a range of factors influencing vector's efficacy in virus transmission, leading to high variability in the transmission efficiency within vector populations. Main shortcomings of most studies are the missing information on the intrinsic potential of individual insects to serve as efficient vectors, both at phenotypic and at genotypic levels. Moreover, detailed analysis of vector competence heredity and monitoring the splitting of both genotypes and phenotypes in filial generations has not been reported. In this study, using the model system Frankliniella occidentalis and Tomato spotted wilt virus, we evaluated the inheritance and stability of the trait vector competence in a population through basic crossings of individually characterized partners, as well as virgin reproduction. We hypothesized that the trait is heritable in F. occidentalis and is controlled by a recessive allele. From the results, 83% and 94% of competent and noncompetent males respectively, inherited their status from their mothers. The trait was only expressed when females were homozygous for the corresponding allele. Furthermore, the allele frequencies were different between males and females, and the competent allele had the highest frequency in the population. These suggest that the trait vector competence is inherited in single recessive gene in F. occidentalis, for which the phenotype is determined by the haplodiploid mechanism. These findings are fundamental for our understanding of the temporal and spatial variability within vector populations with respect to the trait vector competence and at the same time offer an essential basis for further molecular studies. 相似文献