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11.
Abstract The genomes of DNA phage ΦX174 and of RNA phage MS2 each encode a single lysis protein, E protein and L protein, respectively. Based on the known DNA and protein sequences, and with the aid of structural predictions of the proteins, a chimeric gene was constructed. The resulting protein was composed of the N-terminal sequence of E protein and the C-terminal sequence of L protein. The truncated E and L polypeptides used in this construction were functionally inactive. However, the chimeric gene product had very high lysis-inducing activity. This construction is an example which could be extended to the engineering of other lysis proteins designed with specific biotechnological processes in mind. 相似文献
12.
Distribution of δ-aminolevulinic acid biosynthetic pathways among phototrophic bacterial groups 总被引:12,自引:0,他引:12
Two biosynthetic pathways are known for the universal tetrapyrrole precursor, -aminolevulinic acid (ALA). In the ALA synthase pathway which was first described in animal and some bacterial cells, the pyridoxal phosphate-dependent enzyme ALA synthase catalyzes condensation of glycine and succinyl-CoA to form ALA with the loss of C-1 of glycine as CO2. In the five-carbon pathway which was first described in plant and algal cells, the carbon skeleton of glutamate is converted intact to ALA in a proposed reaction sequence that requires three enzymes, tRNAGlu, ATP, Mg2+, NADPH, and pyridoxal phosphate. We have examined the distribution of the two ALA biosynthetic pathways among various genera, using cell-free extracts obtained from representative organisms. Evidence for the operation of the five-carbon pathway was obtained by the measurement of RNase-sensitive label incorporation from glutamate into ALA, using 3,4-[3H]glutamate or 1-[14C]glutamate as substrate. ALA synthase activity was indicated by RNase-insensitive incorporation of label from 2-[14C]glycine into ALA. The distribution of the two pathways among the bacteria tested was in general agreement with their previously established phylogenetic relationships and clearly indicates that the five-carbon pathway is the more ancient process, whereas the pathway utilizing ALA synthase probably evolved much later. The five-carbon pathway is apparently the more widely utilized one among bacteria, while the ALA synthase pathway seems to be limited to the subgroup of purple bacteria.Abbreviations ALA
-aminolevulinic acid
- DTT
dithiothreitol
- PALP
pyridoxal phosphate
- SDS
sodium dodecyl sulfate
- tricine
N-tris-(hydroxymethyl)methylglycine 相似文献
13.
The lipopolysaccharides (LPS) of a rough (R) and a smooth (S) strain of Pseudomonas syringae pv. phaseolicola were analysed. The S-LPS revealed markedly more rhamnose and fucose, but less glucose, than the R-LPS. The presence of 3-O-methyl-rhamnose (acofriose) in the S-LPS was confirmed by cochromatography with authentic acofriose. SDS polyacrylamide gel electrophoresis of the S-LPS demonstrated a cluster of regularly spaced high molecular weight fractions, which was almost lacking in the R-LPS. The main fatty acids of the lipid A of both LPS species were 3-OH-10:0,3-OH-12:0,2-OH-12:0, and 12:0. Two N-linked diesters were demonstrated: 3-O(12:0)-12:0 and 3-O(2-OH-12:0)-12:0. S-LPS was subjected to mild hydrolysis and the degraded polysaccharide separated into three fractions by gel permeation chromatography on a Fractogel TSK HW-50 column. Fraction I, representing nearly only the O-specific side chain, consisted of rhamnose and fucose in a molar ratio of 4:1, with 4% of the rhamnose being 3-O-methylated (acofriose). Fraction II, representing mostly core material, was composed of glucose, rhamnose, heptose, glucosamine, galactosamine, alanine, and a still unidentified amino compound, in an approximate molar ratio of 3:1:1:1:1:1:1, and KDO. Fraction III consisted of released monomers and salts. The LPS was highly phosphorylated (3.28% phosphorus in the core fraction). The thus characterized composition of the LPS O-chain seems to be unique for the pathovar phaseolicola of P. syringae, although many similarities exist to other pathovars as well as to other bacterial species.Abbreviations LPS
lipopolysacchairdes
- GC/MS
combined gas liquid chromatography-mass spectrometry
- HVE
high voltage electrophoresis
- KDO
2-keto-3-deoxyoctonic acid
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecylsulfate
P.s. pv. phaseolicola is termed P. phaseolicola in the text 相似文献
14.
Mark L. Paddock Scott H. Rongey Edward C. Abresch George Feher Melvin Y. Okamura 《Photosynthesis research》1988,17(1-2):75-96
Many herbicides that inhibit photosynthesis in plants also inhibit photosynthesis in bacteria. We have isolated three mutants of the photosynthetic bacterium Rhodobacter sphaeroides that were selected for increased resistance to the herbicide terbutryne. All three mutants also showed increased resistance to the known electron transfer inhibitor o-phenanthroline. The primary structures of the mutants were determined by recombinant DNA techniques. All mutations were located on the gene coding for the L-subunit resulting in these changes Ile229 Met, Ser223 Pro and Tyr222 Gly. The mutations of Ser223 is analogous to the mutation of Ser264 in the D1 subunit of photosystem II in green plants, strengthening the functional analogy between D1 and the bacterial L-subunit. The changed amino acids of the mutant strains form part of the binding pocket for the secondary quinone, Q
b
. This is consistent with the idea that the herbicides are competitive inhibitors for the Q
b
binding site. The reaction centers of the mutants were characterized with respect to electron transfer rates, inhibition constants of terbutryne and o-phenanthroline, and binding constants of the quinone UQ0 and the inhibitors. By correlating these results with the three-dimensional structure obtained from x-ray analysis by Allen et al. (1987a, 1987b), the likely positions of o-phenanthroline and terbutryne were deduced. These correspond to the positions deduced by Michel et al. (1986a) for Rhodopseudomonas viridis.Abbreviations ATP
adenosine 5-triphosphate
- Bchl
bacteriochlorophyll
- Bphe
bacteriopheophytin
- bp
basepair
- cyt c2+
reduced form of cytochrome c
- DEAE
diethylami-noethyl
- EDTA
ethylenediamine tetraacetic acid
- Fe2+
non-heme iron atom
- LDAO
lauryl dimethylamine oxide
- Pipes
piperazine-N,N-bis-2-ethane-sulfonic acid
- PSII
photosystem II
- RC
reaction center
- SDS
sodium dodecylsulfate
- Tris
tris(hydroxy-methyl)aminomethane
- UQ0
2,3-dimethoxy-5-methyl benzoquinone
- UQ10
ubiquinone 50 相似文献
15.
Kenji Kato Su-wan Oh Hiroyuki Yamamoto Takayuki Hanazato Ikuko Yasuda Akira Otuki Masayuki Takahashi 《Ecological Research》1992,7(3):267-276
In order to understand the control mechanisms of a large, stable bacterial standing stock, enclosure experiments were conducted
in a eutrophic lake, where both bacterial productivity and grazing pressure were very high. Total bacterial number in the
different enclosures ranged from 1.2 to 2.7×107 cells mL−1 throughout the experiment. The average bacterial cell production rate estimated from a grazer eliminating experiment was
6.3×105 cells mL−1 h−1. Difference in the bacterial cell production rate between shaded and unshaded enclosures was not apparent. Bacteria showed
a reduction in standing stock of only about 25–30% even after the supply of light was cut to 1%. Bacteria in the shaded enclosures
then recovered their production rate in the first 12 days of perturbation. Grazing pressure in the shaded enclosures was not
less than that for the control. Thus, it was considered a control mechanism of bacterial stable standing stock that the bacteria
shifted their organic substrate from extracellular dissolved organic carbon freshly released from phytoplankton to that already
stocked in the water column, though it is not known whether the dominant bacteria were the same. 相似文献
16.
Relative importance of the effect of 2,4-D,glyphosate, and environmental variables on the soil microbial biomass 总被引:2,自引:0,他引:2
Two post-emergence herbicides (glyphosate and 2,4-D) were applied at field application levels to tilled field plots in a mixed cropping area in south-central Alberta. The effects of these chemicals on certain variables associated with microbial biomass and activity were monitored in these plots (as well as corresponding control plots) for 45 days. Glyphosate did not influence any of the microbial variables tested. Addition of 2,4-D significantly influenced all microbial variables investigated but these effects were transient, being detectable only within the first 1–5 days of herbicide addition. The effects of 2,4-D addition on the microbial variables tested, even when significant, were typically small and probably of little ecological consequence especially when spatial and temporal variation in these variables is taken into account. 相似文献
17.
B. Sinha R. M. Singh U. P. Singh 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1991,82(4):399-404
Summary Studies on the genetics of leaf blight caused byAlternaria triticina using generation mean analysis revealed that additive components played a major role, but that dominance components also contributed significantly in controlling the variability for leaf blight resistance in wheat crosses. Furthermore, the additive x additive type of epistasis was predominant in the first three crosses, whereas in the fourth cross additive x dominance (j) and dominance x dominance (1) components of epistasis were most significant. Because of this it may be desirable to follow a simple recurrent selection scheme for higher tolerance, to isolate resistant plants from the segregating populations derived from crosses of parents of diverse origin following the pedigree method of breeding. CPAN-1887 was very tolerant to leaf blight in the present study and should be utilized in hybridization programs to develop leaf-blight-resistant varieties. 相似文献
18.
F. Amijee E.J. Allans R.N. Waterhouse L.A. Glover A.M. Paton 《Biocontrol Science and Technology》1992,2(3):203-214
L‐forms of the halo blight pathogen, Pseudomonas syringae phaseolicola, were maintained in a medium which suppressed cell wall synthesis. These L‐forms, unlike revertants (walled forms derived from unstable L‐forms) and cell walled (parent) organisms, did not elicit a hypersensitive response in tobacco leaves. Association of L‐forms with Phaseolus vulgaris was established by seed imbibition in L‐form suspensions compared with appropriate control treatments (5% mannitol or heat‐killed cells). Seedling emergence and plant growth was not affected by L‐form imbibition. The association was detected by agglutination assays using polyclonal antibody. The L‐form association was localized to the lower shoot tissue and was progressively lost with age of plants. Plants with associated L‐forms had vigour and shoot weights equivalent to controls and showed no disease symptoms. The cell walled form could not be isolated from plants showing positive agglutination. On challenge with the pathogen, plants associated with L‐forms showed significantly less disease symptoms than controls. Stem extracts, from associated plants, were inhibitory to in vitro cultures of both L‐forms and parent forms of Ps. syr. phaseolicola. These results indicate that L‐form associations confer induced systemic resistance to bean plants and might be developed as novel biocontrol systems. 相似文献
19.
Use of a novel plasmid to monitor the fate of a genetically engineered Pseudomonas putida strain 总被引:1,自引:0,他引:1
Plasmid pSI30 was constructed to increase the sensitivity of detection of a genetically engineered micro-organism (GEM) and its recombinant DNA in environmental samples. This broad host-range, mobilizable plasmid contained chlorocatechol (clc) degradative genes, antibiotic resistance genes (ampicillin and kanamycin) and a fragment of eukaryotic DNA. The clc genes encode enzymes that convert 3-chlorocatechol to maleylacetic acid permitting the host, Pseudomonas putida RC-4, to grow on 3-chlorobenzoate. This catabolic phenotype was exploited using enrichment procedures to detect RC-4(pSI30) cells, free-living in the water column or when irreversibly bound to surfaces. The eukaryotic DNA sequence provided a unique target allowing positive identification by DNA:DNA hybridization. Using the eukaryotic DNA sequence as a probe, no transfer of the plasmid to indigenous bacteria was detected. Persistence of RC-4(pSI30) and its ability to multiply upon addition of 3-chlorobenzoate were demonstrated 78 days after its addition to natural freshwater. In flow-through microcosms RC-4(pSI30), undetectable as free-living cells, was found by enrichment as irreversibly bound sessile forms. These experiments revealed the stability of pSI30 and its utility in a 'combination' detection system for tracking the survival of a GEM and its DNA in environmental samples. 相似文献
20.
The potato has tremendous potential as a transgenic crop and is a good model system by which to analyse metabolic regulation
and gene expression. The potato’s difficult genetics, but ease of genetic transformation and its clonal means of propagation
make it ideal for applied agricultural molecular genetics. Thus, the next 4 years promise to put the potato (with a diversity
of transgenic constructs expressed) in the limelight as many of the first transgenic agricultural products enter the marketplace. 相似文献