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61.
Gene structure and expression of a tobacco endochitinase gene in suspension-cultured tobacco cells 总被引:8,自引:0,他引:8
We have isolated and characterized the genomic clone CHN50 corresponding to tobacco basic endochitinase (E.C.3.2.1.14). DNA sequence and blotting analysis reveal that the coding sequence of the gene present on CHN50 is identical to that of the cDNA clone pCHN50 and, moreover, the CHN50 gene has its origin in the progenitor of tobacco, Nicotiana sylvestris. Tobacco basic chitinases are encoded by a small gene family that consists of at least two members, the CHN50 gene and a closely related CHN17 gene which was characterized previously. By northern blot analysis, it is shown that the CHN50 gene is highly expressed in suspension-cultured tobacco cells and the mRNA accumulates at late logarithmic growth phase. To identify cis-DNA elements involved in the expression of the CHN50 gene in suspensioncultured cells, the chimeric gene consisting of 1.1 kb CHN50 5 upstream region fused to the coding sequence of -glucuronidase (GUS) was introduced by electroporation into protoplasts isolated from suspension-cultured tobacco cells. Transient GUS activity was found to be dependent on the growth phase of the cultured cells, from which protoplasts had been prepared. Functional analysis of 5 deletions suggests that the distal region between -788 and -345 contains sequences that potentiate the high-level expression in tobacco protoplasts and the region (-68 to -47) proximal to the TATA box functions as a putative silencer. 相似文献
62.
Y. Umebachi 《Amino acids》1992,2(1-2):181-187
Summary Papiliochrome II is a pale yellow pigment of butterflies and consists of one molecule each ofL-kynurenine and N--alanyldopamine (NBAD). The aromatic amino nitrogen of kynurenine is bonded to the-carbon of NBAD. There are isomers IIa and IIb which show opposite circular dichroism. The-alanine contents of IIa and IIb were determined and the molar ratio of IIa to IIb has proved to be 1.17. The IIa and IIb were decomposed toL-kynurenine and N--alanylnorepinephrine (NBANE) by being heated in water at 80°C for 30 min. In both IIa and IIb, circular dichroism of the NBANE showed the same positive peak at 280 nm. The NBANE were further decomposed to-alanine and norepinephrine (NE) by being heated in 1 N HC1 at 100°C for 2 hr. The NE was submitted to enantioseparation and has proved to be a racemic mixture in both cases of IIa and IIb. These results are discussed in the light of the enzymic synthesis of IIa and IIb. 相似文献
63.
A phosphorylated analogue of DSIP at Ser7 has been shown to exist endogenously by immunochemical studies. An enzyme which could phosphorylate DSIP has not yet been identified. In the present study, we examined DSIP as a substrate for in vitro phosphorylation by casein kinase II. DSIP was phosphorylated by the enzyme with apparent Km and Vmax values of 20 mM and 90.9 nmol/min/mg protein, respectively. Both ATP and GTP were utilized as phosphoryl donors. Phosphorylation of DSIP was inhibited by heparin and enhanced by spermine. These results demonstrate that DSIP can serve as a possible substrate for casein kinase II in vitro. 相似文献
64.
J. T. Keltjens J. M. H. Hermans G. J. F. A. Rijsdijk C. Van der Drift G. D. Vogels 《Antonie van Leeuwenhoek》1988,54(3):207-220
F430 is the prosthetic group of the methylcoenzyme M reductase of methanogenic bacteria. The compound isolated from Methanosarcina barkeri appears to be identical to the one obtained from the only distinctly related Methanobacterium thermoautotrophicum. F430 is thermolabile and in the presence of acetonitrile or C10
in4
sup-
two epimerization products are obtained upon heating; in the absence of these compounds F430 is oxidized to 12, 13-didehydro-F430. The latter is stereoselectively reduced under H2 atmosphere to F430 by cell-free extracts of M. barkeri or M. thermoautotrophicum. H2 may be replaced by the reduced methanogenic electron carrier coenzyme F420.Abbreviations CH3S-CoM
methylcoenzyme M, 2-methylthioethanesulfonic acid
- HS-CoM
coenzyme M, 2-mercaptoethanesulfonic acid
- F430
Ni(II) tetrahydro-(12, 13)-corphin with a uroporphinoid (III) ligand skeleton
- 13-epi-F430 and 12,13-di-epi-F430
the 12, 13- and 12, 13-derivatives of F430
- 12, 13-didehydro-F430
F430 oxidized at C-12 and C-13
- coenzyme F420
7,8-didemethyl-8-hydroxy-5-deazaflavin derivative
- coenzyme F420H2
reduced coenzyme F420
- MV+
methylviologen semiquinone
- HPLC
high-performance liquid chromatography 相似文献
65.
Addition of 2-(3-chloro-4-trifluoromethyl)anilino-3,5-dinitrothiophene (ANT2p) to detergent-solubilised Photosystem II (PS II) particles results in the photo-oxidation of carotenoid and inhibition of the steady-state oxygen-evolution rate. It has been proposed that ANT2p may modify the water-splitting reactions by mediating the transfer of reducing equivalents from endogenous electron donors, such as carotenoid, to the S2 and S3 oxidation states of PS II. In this paper we present evidence indicating that ANT2p can interact with PS II at two separate loci. The water-splitting complex is shown to be the primary site of attack by ANT2p, since artificial electron donors, such as 1,5-diphenylcarbazide (DPC), can restore PS II photochemical activity by feeding reducing equivalents directly to the reaction centre. The ANT2p interaction at this site is light-intensity dependent. A second inhibitory site close to the reaction centre P-680 chlorophyll is detected at slightly higher ANT2p concentrations. The inhibition at this site is unaffected either by changes in the actinic light intensity or by the addition of electron donors. The flash-induced oxidation of carotenoid has an ANT2p concentration dependence and an insensitivity to DPC which suggests that it results from the inhibition of the reaction centre and not with that of the water-splitting complex. 相似文献
66.
α1 -Adrenergic Receptor-Mediated Downregulation of Angiotensin II Receptors in Neuronal Cultures 总被引:1,自引:0,他引:1
Previous evidence has suggested that brain catecholamine levels are important in the regulation of central angiotensin II receptors. In the present study, the effects of norepinephrine and 3,4-dihydroxyphenylethylamine (dopamine) on angiotensin II receptor regulation in neuronal cultures from rat hypothalamus and brainstem have been examined. Both catecholamines elicit significant decreases in [125I]angiotensin II-specific binding to neuronal cultures prepared from normotensive rats, effects that are dose dependent and that are maximal within 4-8 h of preincubation. Saturation and Scatchard analyses revealed that the norepinephrine-induced decrease in the binding is due to a decrease in the number of angiotensin II receptors in neuronal cultures, with little effect on the receptor affinity. Norepinephrine has no significant actions on [125I]angiotensin II binding in cultures prepared from spontaneously hypertensive rats. The downregulation of angiotensin II receptors by norepinephrine or dopamine is blocked by alpha 1-adrenergic and not by other adrenergic antagonists, a result suggesting that this effect is initiated at the cell surface involving alpha 1-adrenergic receptors. This is further supported by our data indicating a parallel downregulation of specific alpha 1-adrenergic receptors elicited by norepinephrine. In summary, these results show that norepinephrine and dopamine are able to alter the regulation of neuronal angiotensin II receptors by acting at alpha 1-adrenergic receptors, which is a novel finding. 相似文献
67.
A combined single-turnover flash and 35Cl NMR technique has been used to monitor S-state dependence of Cl− binding to PS-II particles derived from mangrove (Avicennia marina). No detectable high-affinity binding was found to particles in the S0 and S1 states, but binding with an affinity comparable to that which activates O2 evolution was found in the S2 and S3 states. 相似文献
68.
L. P. Jones W. A. Compton 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,70(3):318-321
Summary The use of several S1 individuals to represent an S0 individual permits the use of a Design II mating scheme for plants with only one pistillate flower per plant. Estimates of additive (V
A
) and dominance (V
D
) variance from this mating scheme will be biased upwards, when a small number (10) of individuals of each S1 line are used. This bias can be computed, and the additive and dominance estimates can be corrected. Of particular interest is the observation that the additive genetic variance contributes to bias in estimates of V
D
. When S0 plants are non inbred and their selfedprogeny (S1 lines) are used to represent them in developing families for use in the Design II,
where m1 is the number of individuals used to represent an S1 line in developing half sib-families and m2 is the number of individuals used to represent the S1 line in making up full sib-families. For example, in a 3×3 Design II, with about 10 individuals used to represent each S1 line in each cross, m2 = 10 and m1 = 30. When m1 = m2 = 1,
and
Joint contribution from Department of Agronomy, University of Nebraska 68583, and the S. S. Cameron Laboratory, Werribee, Victoria 3030, Australia. Published as paper No. 7395, Journal Series 相似文献
69.
Conversion of atriopeptin II to atriopeptin I by atrial dipeptidyl carboxy hydrolase 总被引:1,自引:0,他引:1
We are examining the substrate specificity of atrial dipeptidyl carboxyhydrolase, a membrane-bound metallo enzyme that we isolated from bovine atrial tissue homogenates. This enzyme readily removes the dipeptide, Phe-Arg, from Bz-Gly-Ser-Phe-Arg, a stand-in substrate for atriopeptin II, one of several atrial natriuretic factors. We now report that the atrial enzyme cleaves the C-terminal dipeptide, Phe-Arg, from atriopeptin II to form atriopeptin I. The km (pH 7.5) is 25 microM and the ratio of relative Vmax/km as a measure of substrate specificity indicates that atriopeptin II is a 240-fold better substrate than Bz-Gly-His-Leu. Only Phe-Arg was detected as a hydrolysis product, indicating that sequential cleavage of Asn-Ser from atriopeptin II does not occur, and that atriopeptin I is not a substrate. Bz-Gly-Asn-Ser was as good a substrate for the atrial enzyme as Bz-Gly-His-Leu, but Bz-Cys(bzl)-Asn-Ser was not hydrolyzed. This result suggests that the presence of an intact disulfide bond or an S-alkylated residue in the P1 position of a substrate (as in atriopeptin I) prevents hydrolysis by the atrial enzyme. Comparative studies were made with the angiotensin I converting enzyme. Atriopeptin II was not a substrate. The stand-in substrates for atriopeptin I, Bz-Cys(bzl)-Asn-Ser and Bz-Gly-Asn-Ser were barely hydrolyzed, which by itself suggests that atriopeptin I is not a substrate of the angiotensin converting enzyme. Our results strongly suggest that atriopeptin II is converted to atriopeptin I and that hydrolysis is mediated by the atrial enzyme. The angiotensin I converting enzyme plays no role in processing these peptides. We suggest that the atrial enzyme be named atrial peptide convertase. 相似文献
70.
Mesenchymal control of branching pattern in the fetal mouse lung 总被引:1,自引:0,他引:1
The effect of mesenchyme on specialization of respiratory epithelium in the fetal mouse was tested in organ cultures. Heterologous combinations were made between respiratory and non-respiratory lung epithelia and the corresponding mesenchymes. Isolated terminal respiratory buds of fetal mouse lungs were recombined with mesenchyme from chick lung parabronchi, mouse trachea or from the avascular, non-respiratory air sacs of chick lungs. Isolated non-branching chick air sacs were combined with mouse terminal bud mesenchyme or mesenchyme from the respiratory branches of chick lungs. Air sac epithelia branched in a pattern characteristic of the chick lung when combined with chick respiratory mesenchyme and in a pattern characteristic of mouse lung when combined with mouse terminal bud mesenchyme. Mouse terminal bud epithelia did not branch with either mouse tracheal mesenchyme or chick air sac mesenchyme but branched in a chick pattern with chick parabronchial mesenchyme. Electron microscopic examination of the cultures showed that all chick air sac epithelial cultures failed to produce surfactant (lamellar bodies) even when they branched. Control cultures of mouse terminal buds contained large numbers of lamellar bodies; mesenchyme which suppressed branching reduced the number of lamellar bodies to only a few in a small proportion of the cells. Culture medium supplemented with growth factors and hormones increased the number of lamellar bodies in heterologous mouse combinations but did not bring the number to control levels. Supplemented medium had no effect on lamellar body production by chick air sac epithelium. The results indicate that branching pattern is determined by the mesenchyme surrounding the epithelial primordium. However, the capacity to synthesize surfactant is determined by the source of the epithelium; mesenchyme may control the degree of expression but not the absolute presence or absence of the differentiated condition. 相似文献