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21.
Establishment of a Sensitized Immunoblotting Method for Measuring Plant Tubulin Content 总被引:1,自引:0,他引:1
A sensitized immunoblotting method was established for measuringsmall amounts of plant tubulin. The method involves electrophoretictransfer of protein including tubulin from SDS-polyacrylamidegels onto nitrocellulose paper, successive incubation of thenitrocellulose paper with a mouse monoclonal antibody to - orß-tubulin of chicken brain, an antibody to mouse IgGas the second antibody and the radioactive iodinated proteinA, and determination of the radioactivities of the bands onthe nitrocellulose paper thus probed. The radioactivities werelinearly proportional to the amounts of - or ß-tubulinfrom dark-grown Vigna mungo seedlings within a range of 4 to56 ng or of 4 to 32 ng, respectively. This method was used to estimate the tubulin contents of severalplant species using Vigna tubulin as a standard. -Tubulin contentsthus estimated were 25, 9, 19 and 11 µg-equivalents ofVigna tubulin per mg protein for Vigna seedlings, Daucus suspensioncells, Catharanthus suspension cells and Mougeolia cells, respectively.ß-Tubulin contents of Vigna, Daucus, Catharanthusand Mougeotia cells were 29, 10, 13 and 5 µg-equivalentsof Vigna tubulin per mg protein, respectively. (Received August 6, 1985; Accepted December 5, 1985) 相似文献
22.
23.
Yuzuru Matsuda Nobuo Nakanishi Geneva Dickens Gordon Guroff 《Journal of neurochemistry》1986,47(6):1728-1734
Soluble extracts from nerve growth factor (NGF)-stimulated PC12 cells prepared by alkaline lysis show a two- to 10-fold greater ability to phosphorylate the 40S ribosomal protein S6 than do extracts from control cells. The alkaline lysis method yields a preparation of much higher specific activity than does sonication. Half-maximal incorporation of 32P from [32P]ATP into S6 occurred after 4-7 min of NGF treatment. The partially purified NGF-sensitive S6 kinase has a molecular weight of 45,000. It is not inhibited by NaCl, chlorpromazine, or the specific inhibitor of cyclic AMP (cAMP)-dependent protein kinase, nor is it activated by addition of diolein plus phosphatidylserine. Trypsin treatment of either crude extracts or partially purified S6 kinase from control or NGF-treated cells was without effect. These data suggest that the S6 kinase stimulated by NGF is neither cAMP-dependent protein kinase or protein kinase C nor the result of tryptic activation of an inactive proenzyme. Treatment of intact cells with dibutyryl cAMP or 5'-N-ethylcarboxamideadenosine also increases the subsequent cell-free phosphorylation of S6. This observation suggests that cAMP-dependent protein kinase may be involved in the phosphorylation of S6 kinase. 相似文献
24.
Although many estimations have been made electronically of mammalian sperm volume, detailed investigations have not been reported for avian spermatozoa with an electronic counter. In the present study, sizing of spermatozoa of fowls and Muscovy and Pekin drakes was examined using a Coulter counter (model ZB). In our preliminary work on fowl sperm volumes, we found mono- or di-morphic distribution displays that were modified depending on the combination of amplification (AMP) and aperture current (APC). Therefore, methodology to estimate the fowl and drake sperm volume was examined. Dilution of semen had no effect on the dimorphic distribution pattern of the sperm volume. Density-gradient centrifugation did not separate two kind of particles in the semen in either continuous or discontinuous Percoll gradients; therefore, we varied settings of AMP and APC, and found that the most suitable settings for measuring sperm volumes of these birds are 1 for AMP and 8 for APC. With these settings, mean volumes of spermatozoa were 5.1 μm3 for fowls, 5.7 μm3 for Muscovy drakes, and 5.6 μm3 for Pekin drakes. 相似文献
25.
Activated c-raf gene in a rat hepatocellular carcinoma induced by 2-amino-3-methylimidazo[4,5-f]quinoline 总被引:8,自引:0,他引:8
F Ishikawa F Takaku M Ochiai K Hayashi S Hirohashi M Terada S Takayama M Nagao T Sugimura 《Biochemical and biophysical research communications》1985,132(1):186-192
A rat hepatocellular carcinoma, IQ7, induced by 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) gave two transformants of NIH 3T3 cells on DNA mediated gene transfer. One of these transformants was examined further and secondary and tertiary transformants were obtained. The secondary transformant was tumorigenic in nude mice. The activated oncogene in this primary transformant was identified as rat c-raf by Southern blot analysis. 相似文献
26.
H Terada T Inagi T Muramatsu H Nagai 《Physiological chemistry and physics and medical NMR》1985,17(1):101-105
Studies were made on determination of the amount of water in rat skin by 1H NMR spectroscopy. The NMR spectrum obtained depended on how the skin was packed into the NMR tube. Consistent results were obtained by packing the skin into a specially designed NMR cell. By this technique the amount of water in the skin could be measured accurately, and results were comparable with those obtained by differential scanning calorimetry. The water content of the skin of rats was consistently about 20% more in females than in males. 相似文献
27.
S Kanazawa H Terada T Iseki S Iwasa K Okuda H Kondo K Okuda 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1986,183(3):333-338
It is now known that nonphysiological cobalamin analogs exist in the gastrointestinal tract, but their metabolic behavior is unclear. In this study, [57Co]cobinamide was used to study its affinity to hog intrinsic factor-cobalamin (IF-Cbl) receptor which has no species specificity against human IF-Cbl receptor, and its relation to human saliva R binder. Cobinamide was prepared from [57Co]cyanocobalamin and separated by paper chromatography. Human IF-Cbl complex was bound to IF-Cbl receptor but free cyanocobalamin was not. Although R binder-cobinamide was not bound to the IF-Cbl receptor, free cobinamide was bound to the IF-Cbl receptor to a significant extent (about one-half of IF-cyanocobalamin binding to the IF-Cbl receptor). We then investigated the binding of cobinamide to R binder and trypsin-treated R binder. Association constant of cobinamide binding to the IF-Cbl receptor was 1.0 X 10(9) M-1 which was much lower than that of cobinamide binding to trypsin-treated R binder and to untreated R binder. Further study indicated that cobinamide binding to the IF-Cbl receptor was blocked by the addition of R binder and also by trypsin-treated R binder. We conclude that one of the roles of R binder is to prevent binding of free cobalamin analogs to the IF-Cbl receptor in the gut. 相似文献
28.
Nobuo Aoki 《Journal of Protein Chemistry》1986,5(4):269-277
Fibrinolytic system is one of the major proteolytic pathways in vivo and primarily responsible for dissolution of thrombi. Two enzymes are primarily involved in this proteolytic system; plasminogen activator (PA) and plasmin. Plasmin is formed by a limited proteolysis of plasminogen by PA, which is mainly synthesized by and secreted from vascular endothelial cells. This proteolytic process proceeds physiologically only on the surface of fibrin. Thus, initiation and progression of the fibrinolytic process depend on the function of endothelial cells and fibrin formation. Endothelial cells may also synthesize and excrete PA inhibitor (PAI) which inhibits immediately, PA once released. The rates of synthesis and excretion of PA and PAI by endothelial cells are regulated by various factors. Among them, thrombin stimulates the release of PA whereas activated protein C may decrease the release of PAI. Thus, both enzymes enhance fibrinolytic potential. PA which has escaped from inhibition by PAI binds to fibrin. 2-Plasmin inhibitor (2PI) inhibits the binding of plasminogen to fibrin, thereby suppressing this fibrin-associated plasminogen activation. A part of 2PI is cross-linked to fibrin by activated factor XIII when fibrin is formed, and the 2PI thus cross-linked to fibrin inhibits in situ plasmin formed on fibrin. Thus, 2PI as well as PAI plays a central role in inhibition of fibrinolysis. 相似文献
29.
Ikuo Taniguchi Hideto Niwa Donald Wong Nobuo Suga 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1986,159(3):331-337
Summary For echolocation, the mustached bat,Pteronotus parnellii rubiginosus, emits orientation sounds (pulses) and listens to echoes. Each pulse is made up of 8 components, of which 4 are constant frequencies (CF1–4) and 4 are frequency-modulated (FM1–4). Target-range information, conveyed by the time delay of the echo FM from the pulse FM, is processed in this species by specialized neurons in a part of the auditory cortex known as the FM-FM area. These cortical neurons are responsive to pulse-echo pairs at specific echo delays (Fig. 1). The essential components in the sound pair include the pulse FM1 followed by an echo FMn (n=2, 3 or 4). Downward sweeping FM1-FMn sounds that are similar to those the animal naturally hears during echolocation are the most effective in evoking facilitative responses. Most FM-FM neurons, however, still exhibit facilitative responses to stimulus pairs consisting of upward sweeping FM sounds and/or pure tones at frequencies found in FM sweeps (Figs. 2 and 3). The magnitude of facilitation is altered by changes in echo rather than pulse amplitude (Figs. 5 and 6). Neurons characterized by shorter best delays (or echoes from closer targets) do not require larger best echo amplitudes for facilitation.Abbreviations
CF
constant frequency
-
FM
frequency modulation
-
H
n
CF — FM harmonics of the mustached bat biosonar signal
-
CF
n
CF components of the harmonics
-
FM
n
FM components of the harmonics
-
PCF
n
pulse CFn
-
ECF
n
echo CFn
-
PFM
n
pulse FMn
-
EFM
n
echo FMn
-
PH
n
pulse Hn
-
EH
n
echo Hn
-
BA
best amplitude for facilitation
-
BD
best delay for facilitation
-
PST
peri-stimulus-time
-
PSTC
peri-stimulus-time-cumulative
-
dB SPL
dB re 20 Pa 相似文献
30.
Temperature dependencies of sprouting and germination were compared for subterranean perennating organs and seeds of ten closely
related species of the genusDioscorea (Dioscoreaceae), a group of monocotyledonous summer perennials which are distributed from the tropics to the subarctic. The
species used wereD. nipponica Makino,D. tokoro Makino,D. japonica Thunb.,D. tenuipes Franch. et Savat.,D. septemloba Thunb.,D. quinqueloba Thunb.,D. izuensis Akahori,D. bulbifera L. f.spontanea (Makino) Makino et Nemoto,D. pentaphylla. L. andD. alata L.; they are distributed from cold northern areas to warmer southern areas approximately in this order in and around Japan.
Bulbil sprouting was also studied in those forming bulbils.
Subterranean organs of the tropical species sprouted faster without any prior temperature treatment, whereas those of species
from the more northern areas sprouted after prechilling. Northern species required longer, periods of prechilling for sprouting.
On the other hand, with seeds or bulbils, the southern species required longer periods of prior temperature treatment for
dormancy breaking. This difference in the length of dormant periods between seeds or bulbils and subterranean organs among
the ten species may be related to their size and position of shedding; seeds or bulbils are small and are shed on the ground
surface, whereas subterranean organs are large and are located below the surface.
It is important to determine in other perennials whether the above relation between dormant features of seeds or bulbils and
subterranean organs are common properties or not. 相似文献