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41.
Expression vectors for cDNA of the κ and λ1 chains of a monoclonal antibody directed against creatine kinase were introduced into murine myeloma cells. κ and γ1 cDNA were either under the control of the SV40 early promoter or of the cognate promoters and enhancers of the light- and heavy-chain genes. Secretion of immuno-reactive κ and γ1 chains into the culture medium was demonstrated with the SV40 promoter as well as with the cognate promoters. Expression of y 1 cDNA with the SV40 early promoter was about twice as high as with the heavy-chain promoter and enhancer. Expression of κ cDNA under the control of the S V40 early promoter was about 17 times higher than with the light-chain promoter and enhancer. These expression levels were compared to those of a genomic immunoglobulin (Ig) κ determinant, including introns. Such an entire κ gene led to expression of the light chain at levels double those with the κ cDNA construction using the SV40 promoter and about 35 times as high when using κ cDNA and the cognate promoter and enhancer. This result might indicate that, besides the cognate promoter and enhancer elements, other intragenic elements are involved in the regulation of Ig expression. However, the SV40 early promoter seems to be able to compensate for the absence of these postulated regulatory elements probably located in the introns. 相似文献
42.
J. Ulrich Ph. U. Heitz T. Fischer E. Obrist F. Gullotta 《Virchows Archiv. B, Cell pathology including molecular pathology》1987,53(1):52-57
Eighteen granular cell tumors from various sites were examined with antisera directed against protein S-100, neuron specific enolase (NSE), alpha-1-antichymotrypsin, and alpha-1-antitrypsin, glial fibrillary acidic protein (GFAP), lysozyme, factor VIII-related antigen, myoglobin and vimentin, as well as with a monoclonal antibody (lu-5) directed against a panepithelial marker. The immunocytochemical reaction pattern of the tumors was heterogeneous. The brain and pituitary tumors and one thyroid tumor reacted for alpha-1-antichymotrypsin and alpha-1-antitrypsin, but not for S-100 protein and NSE. However, tumors from other sites showed immunoreactions for S-100 protein and NSE and some also for vimentin. Reactions for alpha-1-antichymotrypsin and alpha-1-antitrypsin were not observed. All other reactions were similarly negative. We conclude that the morphologically homogeneous group of granular cell tumors is biologically heterogeneous. 相似文献
43.
44.
Photoautotrophic cell suspension cultures of Chenopodium rubrumrequire high concentrations of nitrate and ammonium. Duringthe growth phase total NH4+ and the greater portion of NH3were consumed. During the stationary phase nitrate uptake continuedbut at a substantially smaller rate than during the growth phase.During growth the bulk of the absorbed N was incorporated intoprotein, the amount of which was then maintained constant untilsenescence. NH3 was accumulated upon transition betweenthe growth and the stationary phase. NH3, like the freeamino acids, was deposited in the vacuole but, unlike thesecompounds, could not be remobilized upon transfer of the cellsinto N-free medium. Readdition of NH4+ to the medium, however,resulted in a mobilization of the vacuolar NH3-pool.Reutilization of both vacuolar N-storage pools must have beenaccomplished by recycling from the vacuole to the cytoplasmbecause N-metabolizing enzymes could not be detected in isolatedvacuoles. Transfer of the cells of the stationary phase intomedium containing NH3 and NH4+ resulted in an inductionof nitrate uptake by the cells, but only after a lag phase of45 days. It is conceivable that NH4+ induces NH3-translocatingsystems in the plasmalemma and in the tonoplast. (Received December 19, 1988; Accepted March 2, 1989) 相似文献
45.
46.
Christian Schütt 《Microbial ecology》1989,17(1):49-62
Sixty-two aerobic bacterial strains isolated from the unproductive dystrophic Lake Skärshultsjön (South Sweden) were screened for plasmids. The lake is considered to be an extreme environment because of its high concentration of persistent but nontoxic humic compounds. One-third of the isolates harbored multiple plasmids usually of similar high molecular weights (>25 Mdal). The plasmid-bearing strains were members of the common aquatic taxaPseudomonas spp.,Acinetobacter sp.,Alcaligenes sp.,Aeromonas/Vibrio group, andEnterobacteriaceae (taxonomy is tentative). The majority of isolates displayed multiple resistance to antibiotics and heavy metals. Some of them were capable of degrading aromatic compounds. Three isolates were chosen for curing experiments. Only strain S-68, anAlcaligenes sp., could be cured of one of its two plasmids. It harbored the two cryptic plasmids pQQ32 and pQQ70 of 32 and ca. 70 Mdal, and the latter was segregated during ethidium bromide treatment. Parental strain S-68 was capable of degrading some of nonchlorinated phenolic compounds and displayed resistance to a broad spectrum of antibiotics and the heavy metals Co2+, Ni2+, Zn2+, Cd2+, and Hg2+. Derivative strain S-68-41 lost its resistance to nickel, suggesting segregated plasmid PQQ70 coded for nickel resistance. Transformation experiments to restore nickel resistance in the cured derivative strain were not successful. 相似文献
47.
Schubert Hendrik; Schiewer Ulrich; Tschirner Erhard 《Journal of plankton research》1989,11(2):353-359
The fluorescence characteristics of the cyanobacteria Synechocystisaquatilis Sauv., Microcystis firma (Breb. et Lenorm.) Schmidleand Synechococcus leopoliensis (Racib.) Kom. and the green algaScenedesmus quadricauda (Turp.) Breb. were examined. In thethree cyanobacteria, phycocyanin is the main accessory pigment.Phycoerythrin is not present in our investigated strains ofcyanobacteria. The highest excitation of the chlorophyll a (Chla) fluorescence of cyanobacteria resulted from light with wavelengthsof 620630 nm. A definite Kautsky effectis also evident at this wavelength. However, excitation withblue light (420520 nm) produced only very slight fluorescence.The Kautsky effect is not evident at these wavelengths, evenat high photon flux densities. For Scenedesmus, fluorescencecharacteristics typical of green algae were found. The fluorescenceexcitation of cyanobacteria at 620 nm corresponds to a photosynthesispeak in the action spectrum measured in terms of O2 production.The results underline the necessity of fluorescence measurementsat several wavelengths whenever mixed populations are involved.Such measurements also present possibilities for more accurateestimation of biomass and potential photosynthetic productionin mixed populations. 相似文献
48.
49.
Ursula Rinas Heinrich-Andreas Kracke-Helm Karl Schügerl 《Applied microbiology and biotechnology》1989,31(2):163-167
Summary Glucose supplements to complex growth media of Escherichia coli affect the production of a recombinant model protein under the control of a temperature-sensitive expression system. The bacterial Crabtree effect, which occurs in the presence of glucose under aerobic conditions, not only represses the formation of citric acid cycle enzymes, but also represses the formation of the plasmid-encoded product even though the synthesis of this protein is under the control of the temperature-inducible lambda P
R-promoter/cl857-repressor expression system. When the recombinant E. coli is grown at a moderate temperature (35° C) with protein hydrolysate and glucose as substrates, a biphasic growth and production pattern is observed. In the first phase, the cells grow with a high specific growth rate, utilizing glucose and forming glutamate as a byproduct. The intracellular level of recombinant protein is very low in this phase. Later, glutamate is consumed, indicating an active citric acid cycle. The degradation of glutamate is accompanied by the intracellular accumulation of high amounts of recombinant protein. 相似文献
50.
Glucocorticoid receptor binds cooperatively to adjacent recognition sites. 总被引:22,自引:5,他引:17
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In order to define the mechanism of synergistic induction mediated by multiple glucocorticoid response elements (GRE), the affinity of the glucocorticoid receptor to a single or duplicated GRE was analyzed by gel retardation, nitrocellulose filter binding and by footprinting experiments. Direct measurement of the relative affinity and indirect determination by competition showed greater than 10-fold higher affinity of the glucocorticoid receptor to a duplicated GRE when compared to a single element. Maximal stability of the GRE-receptor complex was obtained using two closely spaced GREs positioned on the same side of the DNA helix. Increasing the distance or changing the helical position of the GREs considerably increased the off rate of the receptor. DNase I footprinting shows in addition to the protection of the GRE region, an altered pattern in the nonprotected intervening DNA indicating structural alteration of the DNA helix by the receptor bound to adjacent GREs. 相似文献