我国若干地区蜡样芽孢杆菌723株噬菌体分型的初步报告

四年来,作者由水中获取蜡样芽孢杆菌噬菌体13株,旨在试作噬菌体分型。待检菌株共723株,由国内16个地区提供(其中食物中毒株121株;食品株602株)。分型结果显示:C29、C30、C24、C27、C19和A_4等型别占优势。中毒株     

Experimental formulations ofBacillus sphaericus for the control of anopheline and culicine larvae

Summary Four experimental formulations ofBacillus sphaericus Neide (2362 isolate) were evaluated for larvicidal activity against culicine and anopheline larvae in several natural and artificial habitats. A granular formulation (5% primary powder) was tested against natural populations of mosquitoes in two simulated habitats in Florida and in maturing and reflooded rice fields in Louisiana. Larvae ofCulex quinquefasciatus Say were reduced by 97 and 99% after application of the granules at the rate of 10 kg/ha to polluted tanks and 2.5 kg/ha to sod-lined potholes, respectively. Anopheline andPsorophora columbiae (Dyar and Knab) larvae were reduced by 68 and 92–100%, respectively, after application of 5 kg granules/ha to rice fields. A flowable concentrate (12.8% primary powder) applied to unpolluted and organically enriched habitats in Florida at 0.25 kg/ha reduced populations ofCulex spp. by 93–100% and 99%, respectively. Sustained-release briquets (5% primary powder) applied at the rate of one half briquet/1.8 m² sod-lined potholes reduced larval populations ofCx. quinquefasciatus by 88–95% for up to 2 weeks in open sunlight. Sustained-release pellets (30% primary powder) applied to small woodland pools in Memphis, TN at the rate of four pellets/pool virtually eliminated larval populations ofCx. restuans Theobald for over 8 days. Variable persistance of larvicidal activity was noted for the other treatments depending on the formulation, target species and habitat.     

Évaluation de la toxicité deBacillus thuringiensis surSpodoptera frugiperda

Résumé Cinquante deux souches deB. thuringiensis appartenant à 13 sérovars ont été testées sur des chenilles néonates deSpodoptera frugiperda en contaminant la surface du milieu semi-synthétique d'élevage. Deux souches du sérovarkenyae et une autre du sérovartolworthi provoquent le plus de mortalité, suivies par les souches des sérovarsaizawai etkurstaki. Les souches les moins actives appartiennent aux sérotypesalesti, dendrolimus, sotto etcolmeri. L'action des souches sur le développement larvaire a aussi été abordée. Les souches des sérovarskenyae, aizawai etkurstaki ont ralenti le développement des chenilles, tandis que les souches des sérovarsalesti, sotto etcolmeri n'ont eu aucun effet.      

The emergence of transgenic potatoes as commercial products and tools for basic science

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.     

Prochymosin expression in Bacillus subtilis

Prochymosin (PC) sequence was cloned in Bacillus subtilis using two kinds of plasmid constructions. In plasmid pSM316 the cDNA was inserted to obtain the intracellular expression of the enzyme. The enzyme turned out to be expressed in an insoluble form which could be converted to native enzyme under proper denaturing and refolding conditions. The levels of intracellular expression of PC were further enhanced by modifying the 5' region of the gene in a way that a two-cistron expression system was created. For the PC secretion, the cDNA was fused to the subtilisin leader sequence and expressed under the control of the B. subtilis neutral protease promoter. A properly folded PC was secreted by the cells, although to low levels.     

Identification of Tn4430, a transposon of Bacillus thuringiensis functional in Escherichia coli

Summary The mobile genetic element Tn4430, originating from the gram-positive bacterium, Bacillus thuringiensis, and previously described as the Th-sequence, is the first transposon isolated from the genus Bacillus. In the present work a gene (APH-III) conferring resistance to kanamycin was inserted into this 4.2 kb transposon. Transposition experiments showed that Tn4430APH-III could transpose in the gram-negative host Escherichia coli when its insertion functions were supplied by an intact copy of Tn4430. By transposing Tn4430APH-III directly onto pBR322, it was possible to determine the nucleotide sequence of the terminal inverted repeats of Tn4430 and of the target DNA site. Identical 38 bp in inverted orientation are situated at each end of the transposon and there is a direct duplication of 5 bp at the insertion site. Thus, it is clear that Tn4430 is closely related to the transposons belonging to the Tn3 family (class II elements).     

Isolation of spore-forming bacilli from mosquitoes in natural breeding habitats in Iraq

New isolates of spore-forming bacilli from larvae and pupae of 3 species of mosquitoes are recorded in central Iraq.Bacillus sphaericus Meyer and Neide was isolated fromCulex pipiens (L.) larva.Bacillus carotarum Koch andBacillus cereus Frankland & Frankland were isolated fromTheobaldia longiareolata (Macquart) pupae.Bacillus laterosporus Laubach andBacillus thuringiensis (H 18) were isolated fromAedes caspius (Pallas) larvae. In addition, unidentifiedBacillus spp. were isolated fromCx. pipiens, T. longiareolata andAe. caspius larvae. Examination of soil samples collected from mosquito natural breeding habitats revealed isolates ofB. cereus, Bacillus thuringiensis H4a 4b; H 12 and H 16 and an unidentifiedBacillus sp.

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Résumé Des souches bactériennes sporogènes sont isolées de moustiques qui se trouvent dans la région centrale de l'Irak. Les résultats obtenus sont les suivants:Bacillus sphaericus Meyer & Neide [Culex pipiens (L.), larve],Bacillus carotarum Koch etBacillus cereus Frankland & Frankland [Theobaldia longiareolata (Macquart), nymphe],Bacillus laterosporus Laubach etBacillus thuringiensis (H 18) [Aedes caspius (Pallas), larvel]. L'examen des larves deCulex pipiens. T. longiareolata etAe. caspius, ainsi que l'analyse des échantillons du sol prélevés dans la région montrent la présence deBacillus cereus, Bacillus thuringiensis H4a 4b; H12 plus H16 et d'autresBacillus non identifiés.

     

Purification and characterization of thermostable leucine dehydrogenase from Bacillus stearothermophilus

Leucine dehydrogenase (l-leucine: NAD⁺ oxidoreductase, deaminating, EC 1.4.1.9) has been purified to homogeneity from a moderate thermophilic bacterium, Bacillus stearothermophilus. Am improved method of preparative slab gel electrophoresis was used effectively to purify it. The enzyme has a molecular mass of about 300,000 and consists of six subunits with identical molecular mass (Mr, 49,000). The enzyme does not lose its activity by heat treatment at 70° C for 20 min, and incubation in the pH range of 5.5–10.0 at 55° C for 5 min. It is stable in 10 mM phosphate buffer (pH 7.2) containing 0.01% 2-mercaptoethanol at over 1 month, and is resistant to detergent and ethanol treatment. The enzyme catalyzes the oxidative deamination of branched-chain l-amino acids and the reductive amination of their keto analogs in the presence of NAD⁺ and NADH, respectively, as the coenzymes. The pH optima are 11 for the deamination of l-leucine, and 9.7 and 8.8 for the amination of -ketoisocaproate and -ketoisovalerate, respectively. The Michaelis constants were determined: 4.4 mM for l-leucine, 3.3 mM for l-valine, 1.4 mM for l-isoleucine and 0.49 mM for NAD⁺ in the oxidative deamination. The B. stearothermophilus enzyme shows similar catalytic properties, but higher activities than that from Bacillus sphaericus.Dedicated to Prof. Dr. G. Drews on the occasion of his 60th birthday     

Exceptionally high copy numbers of a staphylococcal plasmid in Bacillus subtilis revealed by a sandwich hybridization technique

Abstract The copy number of a pUB110 derivative, pKTH10, containing the α-amylase gene from Bacillus amyloliquefaciens , was determined, using an assay based on a sandwich hybridization technique. In this method, a known gene on the plasmid is hybridized between two non-overlapping fragments of that same gene, cloned into separate vectors. One fragment is used as a radiolabelled probe and the other bound to a filter, forming a three-component, 'sandwich' hybrid when the relevant gene is present in the sample. Since the hybridization can only take place in the presence of the relevant gene, the amount of radioactivity binding to the filters will be proportional to the concentration of this gene in the sample. We utilized the α-amylase gene on the plasmid to form the sandwich hybrid. The copy number was of a totally different magnitude from what has previously been reported, and ranged from 2500 copies/viable cell in early logrithimic growth phase to about 500 in late stationary phase.     

The effects of alterations in the suspending medium on low temperature activation of spores of Bacillus stearothermophilus NGB101

The effects of eight different sodium salts on the activation of spores of Bacillus stearothermophilus NGB101 at 30°C were examined. Sodium nitrite was a potent activator spores of NGB101. Complete activation of spore populations was obtained after 6 h or less at 30°C. Activation of spores of NGB101 in solutions of sodium nitrite, like activation in distilled water, was temperature dependent, with optimal activation at 30°C. While a potent activator of spores of NGB101 at 30°C, sodium nitrite was ineffective as an initiator of germination at 65°C. Activation of spores of NGB101 produced marked increases in colony forming spores compared with nonactivated populations. Spore populations activated in solutions of sodium nitrite gave higher plate counts compared with spores activated in distilled water.