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101.
The origin of replication of the IncW plasmid pSa has been cloned and the function of this origin in Escherichia coli examined. A 1.9-kb region of DNA is required for efficient autonomous replication, and a 0.47-kb fragment within this region can initiate replication only in the presence of an autonomously replicating derivative of pSa. An Mr 35,000 protein (repA) is encoded adjacent to the origin and is required for efficient initiation of replication. The derivatives examined provide information suggesting a direct role of partition factors in plasmid replication and incompatibility. 相似文献
102.
R. Vunsh D. Aviv E. Galun 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1982,64(1):51-58
Summary Protoplasts were derived from haploid and diploid Nicotiana sylvestris and N. tabacum. Exposure of the protoplasts to mutagenic doses of ultraviolet (U.V.) radiation prior to two selection rounds in the presence of 4 mM (or 5 mM) and 8 mM of valine, respectively, was required to obtain cell lines with persistent valine resistance. Such lines were obtained from haploid and diploid N. sylvestris protoplasts as well as from haploid protoplasts of N. tabacum but not from (1.8 × 107) diploid N. tabacum protoplasts. The ratio between number of verified valine-resistant cell lines and the initial number of U.V. exposed protoplasts enabled the estimation of the following order of mutation frequency: haploid N. sylvestris > haploid N. tabacum > diploid N. sylvestris. Plants which retained the valine resistance and transmitted it to their sexual progeny were derived from the resistant cell lines. 相似文献
103.
Serotypes of Bacillus thuringiensis and the commercial strain of Bacillus popilliae were examined for their inherent resistance to antibiotics and their mutation potential in respect to neomycin and kanamycin, the presence of which would preclude the use of plasmids marked by genes for resistance to the antibiotics. Clones on initial plates were detected by the recurrence of resistance colonies at superimposable sites on serial replicaplates containing the antibiotic. Susceptible strains were selected for the determination of their antibiotic-resistance mutation potential. Three varieties of B. thuringiensis were found to be doubly resistant, seven varieties were singly resistant (Neor), and three other varieties, including B. popilliae, were susceptible to both antibiotics. Estimates of mutation ratios revealed that three serotypes developed no resistant mutants to either antibiotics in populations as high as 3.0 × 1010; seven other serotypes developed no resistance to kanamycin in populations as high as 4.6 × 109 cells. Three other serotypes exhibited mutation ratios as high as 1.6 × 10?2. We were unable to determine the mutation ratio for B. popilliae. 相似文献
104.
James S. Hutchison Kivie Moldave 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1982,696(1):94-101
A temperature-sensitive mutant of Chinese hamster ovary cells with an altered leucyl-tRNA synthetase fails to grow and to incorporate amino acids into protein properly at or near the non-permissive temperature. This mutant was used to determine whether cessation of growth at the elevated temperature affected elongation factor EF-1, since the activity of EF-1 is markedly lower in non-growing cells in stationary phase than in rapidly-growing cells in exponential phase. Cell-free extracts prepared from cells maintained at 39°C for 24 h showed a marked decrease in the ability to translate natural mRNAs, compared to cells incubated at 34°C. However, the ability to translate poly(U), which requires elongation factor EF-1 (and EF-2), was not affected. Analyses of activities involved in the initiation of protein synthesis and in the activation of amino acids revealed that, with the exception of leucyl-tRNA synthetase, the rest of the components required for translation also appeared to be relatively stable even after 24 h at the elevated temperature. The effects of elevated temperature on cell-free extracts were also investigated. The results were similar to those obtained with intact cells; that is, except for leucyl-tRNA synthetase which was rapidly inactivated in vitro at 39°C, other aminoacyl-tRNA synthetases and translational components involved in chain initiation and elongation were relatively stable. Thus, no change in EF-1 activity was detected as a result of arrested cell growth, an inherent lability of the elongation factor, or metabolic degradation as a consequence of a rapid turnover rate in the absence of protein synthesis. 相似文献
105.
(1) The for 1.3 mM D-allose uptake and efflux in insulin-stimulated adipocytes is 1.7 ± 0.1 min. In the absence of insulin mediated uptake of D-allose is virtually eliminated and the uptake rate () is near that calculated for nonmediated transport. The kinetic parameters for D-allose zero-trans uptake in insulin-treated cells are Kztoi = 271.3 ± 34.2 mM, Vztoi = 1.15 ± 0.12 mM · s?1. (2) A kinetic analysis of the single-gate transporter (carrier) model interacting with two substrates (or substrate plus inhibitor) is presented. The analysis shows that the heteroexchange rates for two substrates interacting with the transporter are not unique and can be calculated from the kinetic parameters for each sugar acting alone with the transporter. This means that the equations for substrate analogue inhibition of the transport of a low affinity substrate such as D-allose can be simplified. It is shown that for the single gate transporter the Ki for a substrate analogue inhibitor should equal the equilibrium exchange Km for this analogue. (3) Analogues substituted at C-1 show a fused pyranose ring is accepted by the transporter. 1-Deoxy-D-glucose is transported but has low affinity for the transporter. High affinity can be restored by replacing a fluorine in the β-position at C-1. The Ki for ; the Ki for . Replacing the ring oxygen also results in a marked reduction in affinity. The Ki for . (4) A hydroxyl in the gluco configuration at C-2 is not required as 2-deoxy-d-galactose (Ki = 20.75 mM) has a slightly higher affinity than d-galactose (Ki = 24.49 mM). A hydroxyl in the manno configuration at C-2 interferes with transport as d-talose (Ki = 35.4 mM) has a lower affinity than d-galactose. (5) d-Allose (Km = 271.3 mM) and 3-deoxy-d-glucose (Ki = 40.31 mM) have low affinity but high affinity is restored by substituting a fluorine in the gluco configuration at C-3. The Ki for . (6) Analogues modified at C-4 and C-6 do not show large losses in affinity. However, 6-deoxy-d-glucose (Ki = 11.08 mM) has lower affinity than d-glucose and 6-deoxy-d-galactose Ki = 33.97 mM) has lower affinity than d-galactose. Fluorine substitution at C-6 of d-galactose restores high affinity. The Ki for . Removal of the C-5 hydroxymethyl group results in a large affinity loss. The Ki. The Ki for . (7) These results indicate that the important hydrogen bonding positions involved in sugar interaction with the insulin-stimulated adipocytes transporter are the ring oxygen, C-1 and C-3. There may be a weaker hydrogen bond to C-6. Sugar hydroxyls in non-gluco configurations may sterically hinder transport. 相似文献
106.
A chemolithoautotrophic type of metabolism, which was hitherto unknown for purple nonsulfur bacteria, was demonstrated by growth experiments using Rhodopseudomonas capsulata Kb1 and Rhodopseudomonas acidophila 10050. These strains were able to grow in a mineral medium in the dark at the expense of H2, O2, and CO2. A minimum doubling time of 9 h was obtained for R. capsulata under an atmosphere containing less than 15% oxygen; higher oxygen concentrations suppressed autotrophic but not chemoorganotrophic growth. Oxygen sensitivity of chemoautotrophically growing cells of R. acidophila was even more pronounced, whereas cells growing chemotrophically on methanol almost tolerated the oxygen concentration of air. Highest oxygen sensitivity of growth of R. acidophila was observed with formate as substrate. The growth yield of cultures grown semiaerobically in the dark on methanol was 0.23 g dry cell material per g methanol consumed. 相似文献
107.
Summary This article is concerned with the determination of kinetic parameters of the Calvin photosynthesis cycle which is described by seventeen nonlinear ordinary differential equations. It is shown that the task requires dynamic data for several sets of initial conditions. The numerical technique is based upon an algorithm for non-linear optimization and Gear's numerical integration scheme for stiff systems of differential equations. The sensitivity of the parameters to noise in the data is tested with a method adapted from Rosenbrook and Storey. A preliminary set of parameters has been obtained from a preliminary set of experimental data. The numerical methods are then tested with synthetic data derived from these parameters. The mathematical model and the results obtained in the simulation are used as an aid in designing new experiments. 相似文献
108.
109.
Summary Three independently isolated tobacco crown gall strains incited byAgrobacterium tumefaciens C58 required phytohormone (auxin and cytokinin) supplements in the basal medium to grow, at 37°C. Six other tobacco crown
gall strains incited, respectively, byA. tumefaciens IIBV7, B6, CGIC, A6NC, 27 and AT4 expressed, at 37°C, the tumor characteristic of ability to grow in vitro on medium lacking
phytohormones. Nopaline was not detectable in C58 tumors cultured at 37°C, but octopine was produced by B6 tumor tissues incibated
at the elevated temperature. C58 tumor strains kept at 37°C for 1 week or more lost the ability to express tumor characteristics
at 27°C such as tissue morphology, growth on basal medium lacking phytohormones and nopaline production. Heat-treated C58
tissues also differed from the original tumor strain in regeneration ability and phytohormone requirements of explants; i.e.
explants from regenerated, heart-treated C58 tumors required both auxin and cytokinin for growth in vitro. 相似文献
110.
The use of acetylene as a convenient assay substrate for nitrogenase in methane oxidising bacteria is complicated by the observation that it is a potent inhibitor of the methane monooxygenase enzyme in both whole cells and cell-free extracts. If the cells were provided with alternative oxidisable carbon substrates other than methane then nitrogen fixing cells would reduce acetylene to ethylene. Hydrogen gas also served as an oxidisable substrate in the assay. Nitrous oxide, which is reduced by nitrogenase to N2 and H2O, was not an inhibitor of methane monooxygenase function and could be used as a convenient assay substrate for nitrogenase. Reduction of both substrates by whole cells showed similar response to oxygen in the assay system and in this respect Methylococcus resembles other free living nitrogen fixing aerobes. 相似文献