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21.
22.
Three distinct regulatory elements comprise the upstream promoter region of the nopaline synthase gene 总被引:5,自引:0,他引:5
Summary Fine deletion mutants were generated in the upstream control region of the nopaline synthase (nos) promoter to define the position and role of upstream regulatory elements. The results indicated that the 8 bp sequence (CAGAAACC) at -106/-113 and its inverted repeat (GGTTTCTG) at -140/-147 are important for promoter function. The downstream element appears more important than the upstream element since deletion of the former reduced promoter activity more significantly than deletion of the latter. Deletion of the element alone, however, did not abolish promoter function, whereas, deletion of the 10 bp potential Z-DNA-forming (Z) element located between the repeat elements nullified promoter activity. Therefore, it appears that the Z element is an essential upstream regulator and the repeated elements are upstream modulators of the nos promoter. These elements are functionally distinct since alteration of stereospecificity or insertion of short oligonucleotides between the elements did not significantly influence promoter activity. These regulatory elements were unable to function from 200 bp upstream of the CCAAT-TATA box region. 相似文献
23.
The two isoenzymes of NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14), previously identified in root nodules of Phaseolus vulgaris L., have both been shown to be located in root-nodule plastids. The nodule specific NADH-GOGAT II accounts for the majority of the activity in root nodules, and is present almost exclusively in the central tissue of the nodule. However about 20% of NADH-GOGAT I activity is present in the nodule cortex, at about the same specific activity as this isoenzyme is found in the central tissue. Glutamine synthetase (GS; EC 6.3.1.2) occurs predominantly as the polypeptide in the central tissue, whereas in the cortex, the enzyme is represented mainly by the polypeptide. Over 90% of both GS and NADH-GOGAT activities are located in the central tissue of the nodule and GS activity exceeds NADH-GOGAT activity by about twofold in this region. Using the above information, a model for the subcellular location and stoichiometry of nitrogen metabolism in the central tissue of P. vulgaris root nodules is presented.Abbreviations Fd-GOGAT
ferredoxin-dependent glutamate synthase
- GOGAT
glutamate synthase
- GS
glutamine synthetase
- NADH-GOGAT
NADH-dependent glutamate synthase
- IEX-HPLC
ion-exchange high-performance liquid chromatography 相似文献
24.
The chalcone synthase multigene family of Petunia hybrida (V30): differential,light-regulated expression during flower development and UV light induction 总被引:11,自引:0,他引:11
We have analysed the expression of the 8–10 members of the gene family encoding the flavonoid biosynthetic enzyme chalcone synthase (CHS) from Petunia hybrida. During normal plant development only two members of the gene family (CHS-A and CHS-J) are expressed. Their expression is restricted to floral tissues mainly. About 90% of the total CHS mRNA pool is transcribed from CHS-A, wheares CHS-J delivers about 10% in flower corolla, tube and anthers. Expression of CHS-A and CHS-J during flower development is coordinated and (red) light-dependent. In young seedlings and cell suspension cultures expression of CHS-A and CHS-J can be induced with UV light. In addition to CHS-A and CHS-J, expression of another two CHS genes (CHS-B and CHS-G) is induced in young seedlings by UV light, albeit at a low level. In contrast to CHS genes from Leguminoseae, Petunia CHS genes are not inducible by phytopathogen-derived elicitors. Expression of CHS-A and CHS-J is reduced to a similar extent in a regulatory CHS mutant, Petunia hybrida Red Star, suggesting that both genes are regulated by the same trans-acting factors. Comparison of the promoter sequences of CHS-A and CHS-J reveals some striking homologies, which might represent cis-acting regulatory sequences. 相似文献
25.
N. Arfman E. M. Watling W. Clement R. J. van Oosterwijk G. E. de Vries W. Harder M. M. Attwood L. Dijkhuizen 《Archives of microbiology》1989,152(3):280-288
The enzymology of methanol utilization in thermotolerant methylotrophic Bacillus strains was investigated. In all strains an immunologically related NAD-dependent methanol dehydrogenase was involved in the initial oxidation of methanol. In cells of Bacillus sp. C1 grown under methanol-limiting conditions this enzyme constituted a high percentage of total soluble protein. The methanol dehydrogenase from this organism was purified to homogeneity and characterized. In cell-free extracts the enzyme displayed biphasic kinetics towards methanol, with apparent K
m values of 3.8 and 166 mM. Carbon assimilation was by way of the fructose-1,6-bisphosphate aldolase cleavage and transketolase/transaldolase rearrangement variant of the RuMP cycle of formaldehyde fixation. The key enzymes of the RuMP cycle, hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), were present at very high levels of activity. Failure of whole cells to oxidize formate, and the absence of formaldehyde-and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formaldehyde via HPS. A comparison of the levels of methanol dehydrogenase and HPS in cells of Bacillus sp. C1 grown on methanol and glucose suggested that the synthesis of these enzymes is not under coordinate control.Abbreviations RuMP
ribulose monophosphate
- HPS
hexulose-6-phosphate synthase
- HPI
hexulose-6-phosphate isomerase
- MDH
methanol dehydrogenase
- ADH
acohol dehydrogenase
- PQQ
pyrroloquinoline, quinone
- DTT
dithiothreitol
- NBT
nitrobluetetrazolium
- PMS
phenazine methosulphate
- DCPIP
dichlorophenol indophenol 相似文献
26.
Francisco Javier Caballero Isabel Igeño Jacobo Cárdenas Francisco Castillo 《Archives of microbiology》1989,152(5):508-511
The phototrophic bacterium Rhodobacter capsulatus E1F1 assimilates ammonia and other forms of reduced nitrogen either through the GS/GOGAT pathway or by the concerted action of l-alanine dehydrogenase and aminotransferases. These routes are light-independent and very responsive to the carbon and nitrogen sources used for cell growth. GS was most active in cells grown on nitrate or l-glutamate as nitrogen sources, whereas it was heavily adenylylated and siginificantly repressed by ammonium, glycine, l-alanine, l-aspartate, l-asparagine and l-glutamine, under which conditions specific aminotransferases were induced. GOGAT activity was kept at constitutive levels in cells grown on l-amino acids as nitrogen sources except on l-glutamine where it was significantly induced during the early phase of growth. In vitro, GOGAT activity was strongly inhibited by l-tyrosine and NADPH. In cells using l-asparagine or l-aspartate as nitrogen source, a concerted induction of l-aspartate aminotransferase and l-asparaginase was observed. Enzyme level enhancements in response to nitrogen source variation involved de novo protein synthesis and strongly correlated with the cell growth phase.Abbreviations ADH
l-alanine dehydrogenase
- AOAT
l-alanine:2-oxoglutarate aminotransferase
- Asnase
l-asparaginase
- GOAT
Glycine: oxaloacetate aminotransferase
- GOGAT
Glutamate synthase
- GOT
l-aspartate: 2-oxoglutarate aminotransferase
- GS
Glutamine synthetase
- HPLC
High-Pressure Liquid Chromatography
- MOPS
2-(N-morpholino)propanesulfonic acid
- MSX
l-methionine-d,l-sulfoximine 相似文献
27.
Aleš Vančura Ivana Vančurová Jan Kopecký Jaroslav Maršálek Daniel Cikánek Gabriela Basařová Vladimir Křišťan 《Archives of microbiology》1989,151(6):537-540
Synthesis of threonine dehydratase in Streptomyces fradiae was positively influenced by valine and negatively by isoleucine. However, these two amino acids had no effect on the activity of this enzyme. Synthesis of threonine dehydratase in -aminobutyrate resistant mutants of S. fradiae was pronouncedly less sensitive to the positive effect of valine and this change in regulation led to valine overproduction. Synthesis of acetohydroxy acid synthase is regulated in a similar manner to that of threonine dehydratase, however a lower level of expression was detected in -aminobutyrate resistant mutants. And again, no effect of branched-chain amino acids on acetohydroxy acid synthase activity was observed. It follows that in S. fradiae synthesis of threonine dehydratase is the main regulatory mechanism governing production and the mutual ratio of synthesized valine and isoleucine.Abbreviations -AB
-aminobutyrate
- AHAS
acetohydroxy acid synthase
- -KB
-ketobutyrate
- MNNG
N-methyl-N-nitro-N-nitrosoguanidine
- TD
threonine dehydratase
- Trans. B.
transaminase of branched-chain amino acids
- VDH
valine dehydrogenase 相似文献
28.
Cecilia Gotor Eloísa Pajuelo Luís C. Romero Antonio J. Márquez José M. Vega 《Archives of microbiology》1990,153(3):230-234
Polyclonal antiserum specific for ferredoxin-nitrite reductase (EC 1.7.7.1) from the green alga Chlamydomonas reinhardii recognized the nitrite reductase from other green algae, but did not cross-react with the corresponding enzyme from different cyanobacteria or higher plant leaves. An analogous situation was also found for ferredoxin-glutamate synthase (EC 1.4.7.1), using its specific antiserum. Besides, the antibodies raised against C. reinhardii ferredoxin-glutamate synthase were able to inactivate the ferredoxin-dependent activity of nitrite reductase from green algae.These results suggest that there exist similar domains in ferredoxin-nitrite reductases and ferredoxin-glutamate synthases from green algae. In addition, both types of enzymes share common antigenic determinants, probably located at the ferredoxin-binding domain. In spite of their physicochemical resemblances, no apparent antigenic correlation exists between the corresponding enzymes from green algae and those from higher plant leaves or cyanobacteria.Abbreviations Fd
ferredoxin
- GOGAT
glutamate synthase
- MV+
reduced methyl viologen (radical cation)
- NiR
nitrite reductase
- PMSF
phenylmethylsulphonyl fluoride
- SDS
sodium dodecyl sulfate 相似文献
29.
Pierre J. Charest Jiro Hattori Janice DeMoor V. N. Iyer Brian L. Miki 《Plant cell reports》1990,8(11):643-646
Summary Genes coding for the enzyme acetohydroxyacid synthase, often referred to as acetolactate synthase (AHAS, ALS; EC 4.1.3.18), from wild type Arabidopsis thaliana and a sulfonylurea-resistant mutant line GH50 (csrl-1; Haughn et al. 1988) were introduced in Nicotiana tabacum. Both genes were expressed at high levels with the 35S promoter. The csrl-1 gene conferred high levels of resistance to chlorsulfuron whereas the wild type gene did not. As selectable markers, chimaeric AHAS genes yielded transgenic plants on chlorsulfuron but at much lower efficiencies than with a chimaeric neomycin phosphotransferase gene on kanamycin (Sanders et al. 1987). Shoot differentiation from leaf discs was delayed on chlorsulfuron by 4–6 weeks. This study indicated a role for mutant AHAS genes in the genetic manipulation of herbicide resistance in transgenic plants but as selectable markers for plant cells undergoing differentiation no advantage over other genes was perceived. 相似文献
30.
Barbara J. Javor 《Archives of microbiology》1988,149(5):433-440
Seven strains of extremely halophilic bacteria (Halobacterium spp., Halococcus spp., and Haloarcula sp.) fixed CO2 under light and dark conditions. Light enhanced CO2 fixation in Halobacterium halobium but inhibited it in Halobacterium volcanii and Haloarcula strain GN-1. Propionate stimulated 14CO2 incorporation in some strains, but inhibited it in others. Semi-starvation in basal salts plus glycerol induced enhanced CO2 fixation rates. 14CO2 fixation in semi-starved cells was stimulated by NH
4
+
or pyruvate and inhibited by succinate and acetate in most strains. No possible reductant was found. In cell-free extracts of H. halobium, NH
4
+
but not propionate stimulated 14CO2 fixation. No RuBP carboxylase activity was detected. The main 14C-labeled -keto acid detected after a 2-min incubation with 14CO2 and pyruvate was pyruvate. Little or no -ketobutyrate was detected among the early products of propionate-stimulated CO2 fixation. Glycine was the major amino acid synthesized during a 2-min incubation with NH
4
+
, propionate, and 14CO2. Propionate-stimulated CO2 fixation was sensitive to trimethoprim and insensitive to avidin. A novel pathway for non-reductive CO2 fixation involving a glycine synthase reaction with CO2, NH
4
+
, and a methyl carbon derived from the -carbon cleavage of propionate is tentatively proposed.Abbreviations used BBS
buffered basal salts
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- MOPS
3-(N-morpholino)propanesulfonic acid
- DNPH
2,4-dinitrophenylhydrazine
- DNP
dinitrophenyl
- TLC
thin-layer chromatography
- FH4
tetrahydrofolate
This work was supported by National Science Foundation grant PCM-8116330 and Petroleum Research Fund grant PRF 13704-AC2 相似文献