| 1. |
- Lindberg, A Michael, et al.
(författare)
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Genome of Coxsackievirus B3
- 1987
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Ingår i: Virology. - : Elsevier BV. - 0042-6822 .- 1096-0341. ; 156:1, s. 50-63
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Tidskriftsartikel (refereegranskat)abstract
- The entire nucleotide sequence of the coxsackievirus B3 strain Nancy (CB3) genome has been determined from cDNA. The genome is 7396 nucleotides long, and encodes a 2185 amino acid long polyprotein. It exhibits the same gene organization as other enterovirus genomes. A detailed comparison was carried out between the proteins encoded by the CB3 and poliovirus type 1 strain Mahoney (PVI) genomes. The genes encoding the VPg polypeptide and the viral polymerase are the most conserved regions. The structural polypeptides VP1, VP2, and VP3 are less well conserved although proline and tryptophan residues frequently are found in identical positions. The VP1 protein of CB3 shows a particularly limited homology in those regions which have been found to induce neutralizing antibodies against PV1. The 5′ noncoding region of CB3 is closely related to that of PV1, with regard to both length and sequence organization, whereas the 3′ noncoding region of CB3 exhibits some unique features.
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| 2. |
- Schnürer, Johan, 1957-, et al.
(författare)
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Effects of moisture on soil microorganisms and nematodes : A field experiment
- 1986
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Ingår i: Microbial Ecology. - : Springer-Verlag New York. - 0095-3628 .- 1432-184X. ; 12:2, s. 217-230
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Tidskriftsartikel (refereegranskat)abstract
- The effects of soil moisture changes on bacteria, fungi, protozoa, and nematodes and changes in oxygen consumption were studied in a field experiment. In one plot the soil was drip-irrigated daily for 10 days, while an adjacent plot experienced one rainfall and was then allowed to dry out. Oxygen consumption was the parameter measured which responded most rapidly to changes in soil moisture content. Lengths of fluorescein diacetate-active hyphae paralleled oxygen consumption in both plots. Total hyphal length was not affected by one rainfall but increased from 700 mg-1 dry weight soil to more than 1,600 m in less than 10 days in the irrigated plot. In the rain plot, bacterial numbers doubled within 3 days and declined during the following period of drought. In the irrigated plot, numbers increased by 50% and then remained constant over the duration of the study. Only small changes in protozoan numbers were observed, with the exception of the last sampling date in the irrigated plot when large numbers of naked amoebae were recorded 2 days after a large natural rainfall. Nematode numbers, especially obligate root feeders, increased in both treatments. The increases were caused by decoiling rather than growth. The results indicate that fungal respiration was dominating, while bacteria, lacking a suitable source of energy, were less active, except for the first days.
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| 3. |
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| 4. |
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| 5. |
- Andersson, Agneta, et al.
(författare)
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Release of aminoacids and inorganic nutrients by heterotrophic marine microflagellates
- 1985
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Ingår i: Marine Ecology Progress Series. - 0171-8630 .- 1616-1599. ; 23, s. 99-106
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Tidskriftsartikel (refereegranskat)abstract
- Heterotrophic microflagellates isolated from the Baltic Sea and grown under laboratoryconditions were shown to release dissolved free amino acids (DFAA) when grazing bacteria. Flagellatesreleased 3H-amino acids when fed 3H-leucine-labelled bacteria, and concentrations of aminoacids increased in the experimental medium. Serine showed a strong positive correlation withflagellate feeding. Aspartic acid, glutamic acid and ornithine also increased more than other aminoacids. During consumption of bacteria, the flagellates released 13% of the ingested nitrogen asammonia, and 30 % of the ingested phosphorus as phosphate. In a field experiment off Scripps Pier, wemeasured bacterial production, flagellate abundance, and concentration of DFAA over a 28 h period.The concentration of DFAA showed a covariation with the flagellate numbers. Results from our fieldand laboratory experiments suggest that flagellates may be a source of DFAA in the sea.
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| 6. |
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| 7. |
- Dahlbäck, B., et al.
(författare)
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Mikrobiologiska aspekter på musselodling
- 1983
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Ingår i: Odling av blåmusslor. - Lund : Signum. - 918533054X ; , s. 60-67
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Bokkapitel (övrigt vetenskapligt/konstnärligt)
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| 8. |
- Fuhrman, J.A., et al.
(författare)
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Diel variations in bacterioplankton, phytoplankton, and related parameters in the Southern California Bight.
- 1985
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Ingår i: Marine Ecology Progress Series. - 0171-8630 .- 1616-1599. ; 27, s. 9-20
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Tidskriftsartikel (refereegranskat)abstract
- The principal objectives of this study were (i) to determine the extent of coupling betweenphytoplankton and microheterotrophs on the shelf off Southern California. (ii) to compare differentmeasures of primary and bacterial secondary production, and (iii) to assess whether sampling timesshould be as strictly controlled for microheterotroph as for autotroph studies. Two diel cycles (May andOctober) were studied by sampling an isotherm as the ship followed paired submerged drogues. Wefound significant die1 changes of chlorophyll, 14C bicarbonate incorporation, bacterial abundance andthymidine incorporation, frequency of dividing bacterial cells (FDC), abundance of non-pigmentedflagellates, particulate organic carbon and nitrogen and their ratios, and dissolved oxygen. Theseparameters all had higher values dunng daylight hours than at night, showing close coupling betweenthe phytoplankton (light-forced) and the microheterotrophs. The ratio of in vivo to extractedchlorophyll a fluorescence, however, displayed a maximum at midnight and minimum at midday,suggesting an endogenous rhythm. Primary production measured by the 14C method was similar to netproduction inferred from in situ oxygen changes. Short-lived peaks in FDC values suggested partlysynchronized bacterial division.
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| 9. |
- Fuhrman, J.A., et al.
(författare)
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Extraction from natural planktonic microorganisms of DNA suitable for molecular biological studies
- 1988
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Ingår i: Applied and Environmental Microbiology. - 0099-2240 .- 1098-5336. ; 54:6, s. 1426-1429
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Tidskriftsartikel (refereegranskat)abstract
- We developed a simple technique for the high-yield extraction of purified DNA from mixed populations ofnatural planktonic marine microbes (primarily bacteria). This is a necessary step for several molecularbiological approaches to the study of microbial communities in nature. The microorganisms from near-shoremarine and brackish water samples, ranging in volume from 8 to 40 liters, were collected on 0.22-,um-pore-sizefluorocarbon-based filters, after prefiltration through glass fiber filters, to remove most of the eucaryotes. DNAwas extracted directly from the filters in 1% sodium dodecyl sulfate that was heated to 95 to 100°C for 1.5 to2 min. This procedure lysed essentially all the bacteria and did not significantly denature the DNA. The DNAwas purified by phenol extraction, and precautions were taken to minimize shearing. Agarose gel electrophoresisshowed that most of the final preparation had a large molecular size (>23 kilobase pairs). The DNA wassufficiently pure to allow complete digestion by the restriction endonuclease Sau3AI and ligation to vector DNA.In a sample in which the extracted DNA was quantified by binding to the dye Hoechst H33258, DNA wasquantitatively extracted, and 45% of the initially extracted DNA was recovered after purification. Final yieldswere a few micrograms of DNA per liter of seawater and were roughly 25 to 50% of the total bacterial DNAin the sample. Alternatives to the initial harvest by filtration method, including continuous-flow centrifugationand thin-channel or hollow-fiber concentration followed by centrifugation, were less efficient than filtration interms of both time and yield, largely because of the difficulty of centrifuging the very small bacteria typical ofmarine plankton. These methods were judged to be less appropriate for studies of natural populations as theyimpose a strong selection for the larger bacteria.
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| 10. |
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| 11. |
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| 12. |
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| 13. |
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| 14. |
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| 15. |
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| 16. |
- Hagström, Åke, et al.
(författare)
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Microbial loop in an oligotrophic pelagic marine ecosystem: Possible roles of cyanobacteria and nanoflagellates in the organic fluxes
- 1988
-
Ingår i: Marine Ecology Progress Series. - 0171-8630 .- 1616-1599. ; 49, s. 171-178
-
Tidskriftsartikel (refereegranskat)abstract
- In an attempt to quantify the organic fluxes within the microbial loop of oligotrophicMediterranean water, organic pools and production rates were monitored. The production of cyanobacteriaand its dynamics dominated the overall productivity in the system. The largest standing stock wasthat of the bacterioplankton and its growth consumed 8.3 pg C 1-' d-', hence about 60 % of the primaryproduction was required for bacterial growth. Using the MiniCap technique, we measured a predationon bacteria of 2 6 X 104 bacteria ml-' h-'. This was in good agreement with the bacterial production rateof 2.3 X 104 cells rnl-' h-' Thus, growth and predation were balanced for heterotrophic bacterioplankton.Almost all of this predation on bacteria was due to organisms passing a 12 vm Nuclepore filter. Thisraises the question of what mechanisms channel 60 % of primary production into bacteria. We thereforeoutlined a mass-balance model to illustrate routes that could explain this transfer. According to ourmodel the main flux route is cyanobacteria and concomitantly consumed heterotrophic bacteria carboninto bacterivores. A substantial fraction of the bacterivore and the microplankton carbon is released byexcretion and/or cell lysis, to be used by the heterotrophic bacterioplankton. About 86% of theautotrophic production is balanced by respiration due to heterotrophic bacteria and protozoa, leaving6 % of the primary production to higher trophic levels. This scenario should apply to ecosystems wherebacterial production rate is high and comparable to primary production, and the dominant primaryproducers are cyanobacteria. A significant fraction of the photosynthetically fixed carbon will bemineralized within a simple microbial loop, thus rendering it an energy sink in the foodweb.
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| 17. |
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| 18. |
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| 19. |
- Horrigan, S.G., et al.
(författare)
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Inorganic nitrogen utilization by assemblages of marine bacteria in seawater culture
- 1988
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Ingår i: Marine Ecology Progress Series. - 0171-8630 .- 1616-1599. ; 50, s. 147-150
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Tidskriftsartikel (refereegranskat)abstract
- Stimulation of heterotrophic bacterial growth by inorganic nitrogen (nitrate and ammonium) was observed in natural assemblages of marine bacteria growth in continuous culture with unsupplemented sea water as primary medium. In the presence of nitrogenous supplements, bacterial numbers increased approximately 3-fold. These results indicate that re-evaluation of the role of heterotrophic bacterioplankton in the pelagic nitrogen cycle may be necessary.
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| 20. |
- Hultmark, Dan, 1949-
(författare)
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Insect immunity : Inducible antibacterial proteins from Hyalophora cecropia
- 1982
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- A powerful bactericidal activity can be induced in the hemolymph of many insects as a response to an injection of bacteria. The nature of the effector molecules of this immune response was investigated, using pupae of the Cecropia moth, Hyalophora cecropia. Three major types of antibacterial proteins were found: the cecropins, the P5 proteins, and lysozyme. They appear in the hemolymph as a result of de novo synthesis.Six different cecropins were purified and characterized. The full amino acid sequences of the three major cecropins A, B and D were determined, as well as partial sequences of the three minor cecropins C, E and F. The cecropins are all very small (Mr = 4,000) and basic (pI > 9.5) proteins, and they show extensive homology in their sequences. The three major cecropins are products of different genes. Their C-terminals are blocked by uncharged groups, which can be removed by mild acid hydrolysis. The minor cecropins are closely related to the major forms, and may be unblocked precursors or, in one case (cecropin F), a minor allelic form. The cecropins were shown to be lytic, and to be efficient against several Gram positive and Gram negative bacterial strains, but not against mammalian cells.The P5 proteins are bactericidal proteins, larger than the cecropins (Mr = 20,000 - 23,000). Six forms, differing in isoelectric point, were isolated. They form two closely related groups, the basic (P5 A-D) and the acidic forms (P5 E-F). Within each group, the different forms have almost identical amino acid compositions.The Cecropia lysozyme is similar to lysozymes isolated from other insects, as well as to that from hen egg white. It is lytic to a restricted number of Gram positive bacteria.The presence of cecropins and other antibacterial factors was demon-strated also in other lepidopterans, notably Galleria mellonella, and may explain earlier observations of antibacterial factors in the latter species.
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| 21. |
- Lacoursière, Jean O., 1958-, et al.
(författare)
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Laboratory study of the influence of water temperature and pH on Bacillus thuringiensis var. israelensis efficacy against black fly larvae (Diptera: Simuliidae)
- 1988
-
Ingår i: Journal of the American Mosquito Control Association. - 8756-971X .- 1943-6270. ; 4:1, s. 64-72
-
Tidskriftsartikel (refereegranskat)abstract
- An experimental formulation of Bacillus thuringiensis var. israelensis was used in the laboratory to assess the influence of water temperature and pH on the relationship between concentration, duration of exposure, and mortality of the northern black fly species Simulium decorum and Prosimulium mixtum/fuscum group. Mortality increases in both species with increases in duration of exposure, concentration, temperature and pH. Onset of death is shortened by increase in concentration and temperature. As temperature rises, the concentration of B. t. i required to induce mortality decreases; the sharpest decline occurring between 12 and 18 degree C for S. decorum , and between 4 and 8 degree C for P. mixtum/fuscum larvae. Lower pH induces a loss of efficacy of the B. t. i. formulation on S. decorum larvae at 4 and 12 degree C.
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| 22. |
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| 23. |
- Norquist, A., et al.
(författare)
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Protection against vibriosis and furunculosis by the use of attenuated strains of Vibrio anguillarum.
- 1989
-
Ingår i: Applied and Environmental Microbiology. - 0099-2240 .- 1098-5336. ; 55:6, s. 1400-1405
-
Tidskriftsartikel (refereegranskat)abstract
- The fish pathogen Vibrio anguillarum causes a lethal infectionin rainbow trout (Salmo gairdneri). Three different avirulentmutants, constructed by transposon insertion mutagenesis (VAN20and VAN70) or as antibiotic-resistant mutants (VAN1000), wereisolated by screening 200 individual isolated mutants for avirulence.When used as live vaccines, all three avirulent mutants wereable to induce protective immunity against the homologous aswell as a heterologous strain of V. anguillarum. When VAN1000was used, protective immunity could be recorded 1 week afterbath vaccination with 10(7) bacteria per ml of water for 30min. A single-dose immunization was effective for at least 12weeks. Western immunoblotting showed that strains of V. anguillarumhave antigenic determinants in common with Aeromonas strains.Therefore, we tested and confirmed that VAN1000 also was ableto induce protective immunity against challenge with Aeromonassalmonicida.
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| 24. |
- Rehnstam, Ann-Sofi, 1959-, et al.
(författare)
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Identification of Vibrio anguillarum in fish by using partial 16S rRNA sequences and a specific 16S rRNA oligonucleotide probe
- 1989
-
Ingår i: Applied and Environmental Microbiology. - 0099-2240 .- 1098-5336. ; 55:8, s. 1907-1910
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Tidskriftsartikel (refereegranskat)abstract
- 16S rRNA from seven different Vibrio anguillarum strains was partially sequenced and compared. From this sequence information we could design a 25-base-long oligonucleotide and use it as a specific probe for identification of V. anguillarum. This was determined by RNA-DNA colony hybridization and slot-blot hybridization. Strong, specific hybridization to the probe was observed for all V. anguillarum strains tested. Furthermore, no cross-hybridization could be seen against five other bacterial species. The detection limit was 5 x 10(3) bacteria per ml. It was even possible to detect V. anguillarum, by slot-blot hybridization, directly in a homogenized kidney from a fish that had died of vibriosis. The partial sequence information revealed small but significant differences between strains of the same species. These sequence differences are sufficiently significant to allow serotyping on the RNA level. Comparing strains of different serotypes revealed a 10-base and an 11-base difference in V. anguillarum serotypes O8 and O9, respectively, in a 122-base partial sequence.
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| 25. |
- Rehnstam, A.-S., et al.
(författare)
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Identification of Vibrio anguillarum in fish using partial 16S rRNA sequences and a specific 16S rRNA oligonucleotide probe.
- 1989
-
Ingår i: Applied and Environmental Microbiology. - 0099-2240 .- 1098-5336. ; 55:8, s. 1907-1910
-
Tidskriftsartikel (refereegranskat)abstract
- 16S rRNA from seven different Vibrio anguillarum strains waspartially sequenced and compared. From this sequence informationwe could design a 25-base-long oligonucleotide and use it asa specific probe for identification of V. anguillarum. Thiswas determined by RNA-DNA colony hybridization and slot-blothybridization. Strong, specific hybridization to the probe wasobserved for all V. anguillarum strains tested. Furthermore,no cross-hybridization could be seen against five other bacterialspecies. The detection limit was 5 x 10(3) bacteria per ml.It was even possible to detect V. anguillarum, by slot-blothybridization, directly in a homogenized kidney from a fishthat had died of vibriosis. The partial sequence informationrevealed small but significant differences between strains ofthe same species. These sequence differences are sufficientlysignificant to allow serotyping on the RNA level. Comparingstrains of different serotypes revealed a 10-base and an 11-basedifference in V. anguillarum serotypes O8 and O9, respectively,in a 122-base partial sequence.
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| 26. |
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| 27. |
- Schnürer, Johan, 1957-, et al.
(författare)
-
Fluorescein diacetate hydrolysis as a measure of total microbial activity in soil and litter
- 1982
-
Ingår i: Applied and Environmental Microbiology. - Washington : American Society for Microbiology. - 0099-2240 .- 1098-5336. ; 43:6, s. 1256-1261
-
Tidskriftsartikel (refereegranskat)abstract
- Spectrophotometric determination of the hydrolysis of fluorescein diacetate (FDA) was shown to be a simple, sensitive, and rapid method for determining microbial activity in soil and litter. FDA hydrolysis was studied in soil and straw incubated for up to 3h. Hydrolysis was found to increase linearly with soil addition. FDA hydrolysis by pure cultures of Fusarium culmorum increased linearly with mycelium addition both in shake cultures and after inoculation into sterile soil. FDA hydrolys is by Pseudomonas denitrificans increased linearly with biomass addition. The FDA hydrolytic activities in soil samples from different layers of an agricultural soil were correlated with respiration. Acetone was found to be suitable for terminating the reaction.
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| 28. |
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| 29. |
- Wikner, Johan, 1961-, et al.
(författare)
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Evidence for a tightly coupled nanoplanktonic predator-prey link regulating the bacterivores in the marine-environment
- 1988
-
Ingår i: Marine Ecology Progress Series. - Oldenburg : Inter-Research. - 0171-8630 .- 1616-1599. ; 50:1-2, s. 137-145
-
Tidskriftsartikel (refereegranskat)abstract
- A coupled predator-prey chain, starting with bactenvores, was invest~gated using the mlnicell recapture technique (MiniCap) Water samples were subjected to slze fract~onation wth decreasing filter pore sue in order to obtain a successive truncation of the microbial food chaln Our results showed that the malor bacterivores were flagellates in the size range of 1 to 3 pm The truncation of the food chain caused increased or decreased predation on the bactena, d e p e n d~n go n whether the bacterivores 'ivere released from or subjected to increased predat~on pressure We present a model describing trophic interactions between organisms less than 12 pm In size This model suggests 4 trophic levels to form a regulatory chain exer t~nga tight control on major bacterivores.
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| 30. |
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| 31. |
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| 32. |
- Öquist, G., et al.
(författare)
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Chlorophyll a fluorescence an alternative method for estimating primary production
- 1982
-
Ingår i: Marine Biology. - 1230-7688. ; 68:1, s. 71-75
-
Tidskriftsartikel (refereegranskat)abstract
- The in vivo chlorophyll a fluorescence index (F+DcM u-F-DcMu/F+DcMU) of natural waters was compared to the 14C-determined primary production, and the fluorescence intensity in the presence of 3-(3,4-dichlorophenyl)-l,l-dimethylurea (F + DCM~) was studied as a function of extracted and spectrophotometrically determined chlorophyll concentrations. Samples were taken every second week from May through October, 1979, at the station "Systrarna" situated in a coastal area of the Bottnian Sea. In addition, samples from the Archipelago Sea of the Baltic were collected on board the Finnish research vessel R/S "Aranda" during the September cruise 1979. The correlations between the fluorescence index and the 14C-determined primary production and between F+DcMu and total chlorophyll concentration were good when samples taken over short time intervals were compared. The shortcomings of both the fluorescence and the 14C methods are discussed. It is concluded that the fluorescence method is useful if it is desirable to follow with high time resolution any changes in the potential for photosynthesis (or pirmary production) in a water mass over relatively short time periods; e.g. during an algal bloom. The fluorescence method can furthermore be technically developed for automatic monitoring with a high time resolution. Efforts are being made in our laboratory to develop the method further to give information about the in situ rates of photosynthesis rather than the potential for photosynthesis in a phytoplankton population.
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| 33. |
- Wold, Agnes E, 1955, et al.
(författare)
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Difference between bacterial and food antigens in mucosal immunogenicity.
- 1989
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Ingår i: Infection and immunity. - 0019-9567. ; 57:9, s. 2666-73
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Tidskriftsartikel (refereegranskat)abstract
- The mucosa-associated lymphoid tissue may deviate from its systemic counterpart in being able to discriminate between microbial and nonmicrobial antigens. To study this, the systemic and mucosal antibody responses to bacterial and food antigens were followed in parallel in female rats during two pregnancies and lactation periods. Germfree rats were monocolonized with an Escherichia coli O6K13H1 strain, and their diet was switched to pellets containing large amounts of ovalbumin and beta-lactoglobulin. Antibodies against O6 lipopolysaccharide already appeared in serum and bile 1 week after colonization, and those against type 1 fimbriae appeared a few weeks later. Serum immunoglobulin G antibodies against the E. coli enzyme beta-galactosidase were found in moderate titers in all rats after 16 weeks of exposure. In contrast, few rats had detectable antibody levels against the dietary proteins ovalbumin and beta-lactoglobulin in serum or bile even after 16 weeks of exposure. In the milk, antibodies against E. coli beta-galactosidase and type 1 fimbriae reached the highest titers, while moderate titers were found against the food antigens and against O6 lipopolysaccharide. The difference in immune reactivity against bacterial versus dietary antigens was not likely due to insufficient amounts of the latter reaching lymphoid tissue, since (i) uptake studies indicated that ovalbumin was more efficiently taken up than endotoxin and (ii) the same difference in antigenicity between ovalbumin and E. coli was seen after immunization directly into Peyer's patches. We therefore suggest that a prerequisite for a strong mucosal antibody response is that the antigen be encountered by the gut-associated lymphoid tissue within microorganisms capable of stimulating antigen presentation.
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| 34. |
- Aevarsson, A, et al.
(författare)
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Ligands to the 2Fe iron-sulfur center in succinate dehydrogenase
- 1988
-
Ingår i: FEBS Letters. - : Wiley. - 1873-3468 .- 0014-5793. ; 232:2, s. 298-302
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Tidskriftsartikel (refereegranskat)abstract
- Membrane-bound succinate oxidoreductases are flavoenzymes containing one each of a 2Fe, a 3Fe and a 4Fe iron-sulfur center. Amino acid sequence homologies indicate that all three centers are located in the Ip (B) subunit. From polypeptide and gene analysis of Bacillus subtillis succinate dehydrogenase-defective mutants combined with earlier EPR spectroscopic data, we show that four conserved cysteine residues in the first half of Ip are the ligands to the [2Fe-2S] center. These four residues have previously been predicted to be the ligands. Our results also suggest that the N-terminal part of B. subtilis Ip constitutes a domain which can incorporate separately the 2Fe center and interact with Fp, the flavin-containing subunit of the dehydrogenase.
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| 35. |
- Carlsson, Peter, et al.
(författare)
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Bacillus subtilis citM, the structural gene for dihydrolipoamide transsuccinylase: cloning and expression in Escherichia coli
- 1987
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Ingår i: Gene. - : Elsevier BV. - 1879-0038 .- 0378-1119. ; 61:2, s. 217-224
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Tidskriftsartikel (refereegranskat)abstract
- The 2-oxoglutarate dehydrogenase multienzyme complex is composed of three different subenzymes: 2-oxoglutarate dehydrogenase (E1o), dihydrolipoamide transsuccinylase (E2o), and dihydrolipoamide dehydrogenase (E3). Bacillus subtilis E1o and E2o are encoded by the citK and citM genes, respectively. A 3.4-kb BamHI DNA fragment containing citK and citM markers was isolated from a library of B. subtilis DNA in Escherichia coli. Functional E2o was expressed from the cloned DNA both in B. subtilis and E. coli. E2o had an apparent Mr of 60000 when expressed in E. coli. The B. subtilis E2o component complemented an E. coli E2o-defective mutant in vivo and in vitro. It is concluded that functional B. subtilis E2o can be produced in E. coli and can interact with E. coli and E1o and E3 to form an active chimeric enzyme complex.
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| 36. |
- Carlsson, P., et al.
(författare)
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Genetic Characterization of Bacillus subtilis odhA and odhB, encoding 2-oxoglutarate dehydrogenase and dihydrolipoamide transsuccinylase, respectively
- 1989
-
Ingår i: Journal of Bacteriology. - : American Society for Microbiology. - 0021-9193 .- 1098-5530. ; 171:7, s. 3667-3672
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Tidskriftsartikel (refereegranskat)abstract
- The 2-oxoglutarate dehydrogenase complex consists of three different subenzymes, the E1o (2-oxoglutarate dehydrogenase) component, the E2o (dihydrolipoyl transsuccinylase) component, and the E3 (dihydrolipoamide dehydrogenase) component. In Bacillus subtilis, the E1o and E2o subenzymes are encoded by odhA and odhB, respectively. A plasmid with a 6.8-kilobase-pair DNA fragment containing odhA and odhB was isolated. Functional E1o and E2o are expressed from this plasmid in Escherichia coli. Antisera generated against B. subtilis E1o and E2o expressed in E. coli reacted with antigens of the same size from B. subtilis. The nucleotide sequence of odhB and the terminal part of odhA was determined. The deduced primary sequence of B. subtilis E2o shows striking similarity to the corresponding E. coli protein, which made it possible to identify the lipoyl-binding lysine residue as well as catalytic histidine and aspartic acid residues. An mRNA of 4.5 kilobases hybridizing to both odhA and odhB probes was detected, indicating that odhA and odhB form an operon.
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| 37. |
- Carlsson, Peter, et al.
(författare)
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In vitro complementation of Bacillus subtilis and Escherichia coli 2-oxoglutarate dehydrogenase complex mutants and genetic mapping of B. subtilis citK and citM mutations
- 1986
-
Ingår i: FEMS Microbiology Letters. - : Oxford University Press (OUP). - 1574-6968 .- 0378-1097. ; 37:3, s. 373-378
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Tidskriftsartikel (refereegranskat)abstract
- AbstractThe 2-oxoglutarate dehydrogenase complex of the tricarboxylic acid cycle (TCA) consists of multiple copies of 3 different subenzymes; E1, E2 and E3. The E3 subenzyme is also a component of the pyruvate dehydrogenase complex. Bacillus subtilis 2-oxoglutarate dehydrogenase mutants were studied. The mutants defective in E1, E2 and E3 subenzyme activity, respectively, could be separated into 3 groups by biochemical complementation analyses. The groups correspond to the citK, citM and citL genes. A B. subtilis subenzyme defect, probably E1, could be complemented with the corresponding Escherichia coli wild-type subenzyme and vice versa. Mutations in citK and citM are closely linked. The gene order kauA--- ---citK-citM was determined from 3-factor transformation crosses. It is concluded that the gene organization and the subenzyme structure of the 2-oxoglutarate dehydrogenase complex are similar in B. subtilis and E. coli.
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| 38. |
- Fridén, H, et al.
(författare)
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Cytochrome b558 of Bacillus subtilis
- 1988
-
Ingår i: Cytochrome Systems : Molecular Biology and Bioenergetics - Molecular Biology and Bioenergetics. - 9781461290780 - 9781461319412 ; , s. 641-647
-
Bokkapitel (refereegranskat)abstract
- The membranebound tricarboxylic acid cycle enzyme succinate dehydrogenase (SDH) is associated with a b-type cytochrome in many organisms. 1,2 The cytochrome b is often found in stoichiometric amounts in isolated succinate-ubiquinone oxidoreductase (complex II) from bovine heart,3Neurospora crassa,4Ascaris suum5 and plant6 mitochondria as well as in SDH complexes isolated from both Gram-negative and Gram-positive8,9 bacteria whereas yeast (Saccharomyces cerevisiae) apparently lacks this type of cytochrome.10
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| 39. |
- Fridén, H, et al.
(författare)
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Genetic and biochemical characterization of Bacillus subtilis mutants defective in expression and function of cytochrome b-558
- 1987
-
Ingår i: European Journal of Biochemistry. - : Wiley. - 0014-2956 .- 1432-1033. ; 168, s. 695-701
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Tidskriftsartikel (refereegranskat)abstract
- Bacillus subtilis succinate dehydrogenase is bound to the cytoplasmic membrane by cytochrome b-558, a 23-kDa transmembrane protein which also functions as electron acceptor to the dehydrogenase. The structural gene for the apocytochrome, sdhC, has previously been cloned and sequenced. In this work the structure and translation of cytochrome b-558 was studied in different sdhC mutants. Mutant cytochrome was analyzed both in B. subtilis and after amplification in Escherichia coli. It is concluded that amino acid substitutions in the C-terminal half of the cytochrome can prevent the binding of succinate dehydrogenase without affecting membrane binding of the cytochrome protein or heme ligation. Mutagenesis of His-113 excludes this residue as an axial heme ligand. A base-pair exchange of G to A in the ribosome-binding sequence of sdhC was found to reduce cytochrome b-558 translation about tenfold in B. subtilis, whereas the mutation had no effect on translation in E. coli. Translation of the two succinate dehydrogenase genes from the sdhCAB polycistronic transcript does not seem to be coupled to translation of sdhC. Less than 10% of the wild-type amount of membrane-bound succinate dehydrogenase in B. subtilis still allows growth on non-fermentable substrate, but makes the dehydrogenase a limiting enzyme in the tricarboxylic acid cycle and leads to succinate accumulation.
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| 40. |
- Hederstedt, Lars, et al.
(författare)
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Covalent binding of FAD to Bacillus subtilis succinate dehydrogenase
- 1987
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Ingår i: Flavins and Flavoproteins : Proceedings of the Ninth International Symposium, Atlanta, Georgia, U. S. A. June 7-12, 1987 - Proceedings of the Ninth International Symposium, Atlanta, Georgia, U. S. A. June 7-12, 1987. - 3110109506 - 0899253067 ; , s. 729-736
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Konferensbidrag (refereegranskat)
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| 41. |
- Hederstedt, Lars, et al.
(författare)
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New properties of Bacillus subtilis succinate dehydrogenase altered at the active site
- 1989
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Ingår i: Biochemical Journal. - : Portland Press Ltd.. - 0264-6021 .- 1470-8728. ; 260:2, s. 491-497
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Tidskriftsartikel (refereegranskat)abstract
- Mammalian and Escherichia coli succinate dehydrogenase (SDH) and E. coli fumarate reductase apparentlycontain an essential cysteine residue at the active site, as shown by substrate-protectable inactivation withthiol-specific reagents. Bacillus subtilis SDH was found to be resistant to this type of reagent and containsan alanine residue at the amino acid position equivalent to the only invariant cysteine in the flavoproteinsubunit of E. coli succinate oxidoreductases. Substitution of this alanine, at position 252 in the flavoprotein subunit of B. subtilis SDH, by cysteine resulted in an enzyme sensitive to thiol-specific reagents and protectable by substrate. Other biochemical properties of the redesigned SDH were similar to those of the wild-type enzyme. It is concluded that the invariant cysteine in the flavoprotein of E. coli succinate oxidoreductases corresponds to the active site thiol. However, this cysteine is most likely not essential for succinate oxidation and seemingly lacks an assignable specific function. An invariant arginine in juxtaposition to Ala-252 in the flavoprotein of B. subtilis SDH, and to the invariant cysteine in the E. coli homologous enzymes, is probably essential for substrate binding.
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| 42. |
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| 43. |
- Petricek, M, et al.
(författare)
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The structural gene for aspartokinase II in Bacillus subtilis is closely linked to the sdh operon
- 1989
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Ingår i: FEMS Microbiology Letters. - 1574-6968. ; 61:1-2, s. 85-87
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Tidskriftsartikel (refereegranskat)abstract
- The aecA and aecB loci map at 250 and 290 degrees, respectively, on the Bacillus subtilis chromosomal genetic map. The aecB locus has been proposed as the structural gene for aspartokinase II. From DNA sequence analyses and comparisons to the sequence of the aspartokinase II gene, it can be concluded that the structural gene for aspartokinase II is located close to sdh at 250 degrees and cannot be aecB. A detailed map over 7 kbp in the 250 degree region is presented.
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| 44. |
- Tham, Wilhelm, 1951-, et al.
(författare)
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A comparison of six media for isolating Staphylococcus aureus from foods
- 1987
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Ingår i: Food microbiology (Print). - : Elsevier. - 0740-0020 .- 1095-9998. ; 4:2, s. 133-146
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Tidskriftsartikel (refereegranskat)abstract
- Six culture media were compared with blood agar regarding colony counts and colony diameters of 53 Staphylococcus aureus strains. There were no statistically significant differences between the media. The forced differentiation of the results via cluster analysis, however, gave some indications that differences existed. In terms of colony counts, LSM, BP Oxoid and BP Oxoid + pp gave results similar to those given by blood agar. Colony diameter seemed to be a doubtful measure of the media's suitability and none of the six media showed similar diameter values to those of blood agar. As regards the appearance of the S. aureus strains on the six media, BP BBL and LSM corresponded the most closely to the inventors descriptions. The selectivities of the media were tested by cultivating 102 food samples from various sources. The most favourable retardation against micro-organisms other than S. aureus was shown by BP BBL, KRANEP and LSM. In terms of all tests performed, BP BBL was the most satisfactory medium for isolating S. aureus. PCVJ was the poorest of all media used in this study.
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| 45. |
- Tham, Wilhelm, 1951-
(författare)
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Histamine formation by enterococci isolated from home-made goat cheeses
- 1988
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Ingår i: International Journal of Food Microbiology. - : Elsevier. - 0168-1605 .- 1879-3460. ; 7:2, s. 103-108
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Tidskriftsartikel (refereegranskat)abstract
- A survey was made of the histamine-producing capability of enterococci isolated from goat cheeses. The strains, 130 Streptococcus faecium and 106 S. faecalis, were grown in Trypticase Soy Broth Histidine medium (TSBH) at 35°C for 24 h and the histamine formed was determined by fluorometry. Forty-one (31.5%) of the S. faecium strains and 2 (1.9%) of the S. faecalis strains produced histamine. The largest amount detected was 4.0 μg histamine/ml TSBH. Compared with the amounts of histamine produced by some Gram-negative bacteria, the histamine production by enterococci seems to be low. Under the present conditions the enterococci seem to have no relevance from a histamine intoxication point of view.
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| 46. |
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| 47. |
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| 48. |
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| 49. |
- Andersson-Nordström, Agneta, 1958-
(författare)
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Flagellates in the marine microbial food web : the ecology of a mixotrophic nanoflagellate, Ochromonas sp.
- 1989
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Nanoflagellates were found to be abundant in a coastal area of the northern Bothnian Sea. The maximum concentration of nanoflagellates, approximately 8000 cells ml-1, was observed in July, coinciding with a decrease in the abundance of cyanobacteria. Pigmented and non-pigmented nanoflagellates were approximately equally distributed throughout the year. Most of the identified genera are known as being phagotrophic, independent if autotrophic or not.A non-cyst-forming pigmented flagellate, Ochromonas sp., was isolated and nutritionally characterized. This chrysophycean flagellate was shown to be a mainly heterotrophic organism: Photosynthesis was too poor to support multiplication of the cells, whereas when feeding on bacteria, high growth rates were obtained. The biological function of the photosynthetic apparatus is suggested to be a survival mechanism during poor bacterial conditions.The flagellate grazed bacteria selectively, preferring cyanobacteria and large cells of heterotrophic bacteria, presumably depending on size-selective grazing. Despite higher growth rates of the bacteria in the sea during summer (July) than spring (May), heterotrophic bacteria in the sea was observed to be smaller in the summer. Nanoflagellates showed a maximum in July, and by selective grazing of large bacteria they might have caused the decrease in the average size of the bacteria and the decrease in the abundance of cyanobacteria.During the consumption of bacteria the flagellate was shown to remineralize nutrients at high rates and excrete dissolved free amino acids. Assuming the existence of a protozoan predator-prey chain of several trophic levels, it seems likely that a significant part of the nutrients fixed by primary producers is remineralized in the euphotic zone. Furthermore, data from this work indicate that flagellate activity may be a significant source of dissolved free amino acids, utilizable for the heterotrophic bacteria.
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| 50. |
- Brandsch, Roderich, et al.
(författare)
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Expression and flavinylation of Arthrobacter oxydans 6-hydroxy-D-nicotine oxidase in Bacillus subtilis
- 1989
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Ingår i: Journal of General Microbiology. - : Microbiology Society. - 0022-1287. ; 135:1093-1099
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Tidskriftsartikel (refereegranskat)abstract
- 6-Hydroxy-d-nicotine oxidase (6-HDNO) of Arthrobacter oxydans, an enzyme inducible by dl-nicotine, contains FAD covalently bound via an 8α-N(3)His linkage. Expression of the gene encoding 6-HDNO and flavinylation of the protein were studied in Bacillus subtilis. In this heterologous system the following findings were made. 1. An enzymically active covalently flavinylated 6-HDNO of normal size can be expressed in B. subtilis. 2. The natural promoter of the 6-HDNO gene appeared inefficient in B. subtilis. The B. subtilis sdh promoter, when inserted upstream of the A. oxydans promoter, increased 6-HDNO expression >50-fold. 3. Expression of the 6-HDNO gene from plasmids in B. subtilis was, independently of the promoter construct used, stimulated more than fivefold by dl-nicotine in the growth medium. It is concluded that flavinylation of 6-HDNO is possibly autocatalytic and mediated by factors generally found in bacterial cells.
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