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71.
Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. 总被引:69,自引:2,他引:67 下载免费PDF全文
Using a set of synthetic oligonucleotides homologous to broadly conserved sequences in-vitro amplification via the polymerase chain reaction followed by direct sequencing results in almost complete nucleotide determination of a gene coding for 16S ribosomal RNA. As a model system the nucleotide sequence of the 16S rRNA gene of M.kansasii was determined and found to be 98.7% homologous to that of M.bovis BCG. This is the first report on a contiguous sequence information of an entire amplified gene spanning 1.5 kb without any subcloning procedures. 相似文献
72.
Toxicity and isolation of the cyanobacterium Nodularia spumigena from the southern Baltic Sea in 1986 总被引:1,自引:1,他引:0
Three water bloom samples were collected in August 1986 from the southern Baltic Sea. Acute toxicity of the samples was determined by mouse bioassay and the toxins were further studied by HPLC. The bloom samples contained equal amounts of cyanobacteria Nodularia spumigena and Aphanizomenon flos-aquae and were hepatotoxic. Two hepatotoxic Nodularia spumigena strains were isolated from the samples. The isolates produce a toxic peak indistinguishable from the bloom material in the HPLC analysis. The toxicity of the fractions was verified by mouse bioassay. Thus the toxicity of the bloom samples was in all likelihood caused by Nodularia spumigena. 相似文献
73.
The 76 kD cell-adhesion factor from crayfish haemocytes promotes encapsulation in vitro 总被引:2,自引:0,他引:2
Summary Semigranular cells from the crayfish, Pacifastacus leniusculus, were separated by Percoll gradient centrifugation and were used to study the encapsulation of foreign particles. The semigranular cells were found strongly to encapsulate glass beads coated with haemocyte lysate in which the prophenoloxidase-activating system had been activated with laminarin or with a low concentration of calcium ions. The granular cells only weakly encapsulated these particles. The encapsulationpromoting factor was purified from haemocyte lysates and found to be a 76 kD protein which was recognized by an antiserum to the previously described 76 kD cell-adhesion factor. After the last step in purification (Con A-Sepharose chromatography), the flowthrough consisted of several proteins, which had some, but less, encapsulation-promoting activity and contained a 30 kD band that was also recognized by the antiserum to the 76 kD cell-adhesion factor. If the haemocyte lysate prepared in low [Ca2+] was incubated with a -1,3-glucan prior to purification, no 76 kD protein could be isolated but only a 30 kD protein. The 30 kD protein thus seems to be a degradation product of the 76 kD cell-adhesion factor. We conclude that the 76 kD protein which is released from degranulating haemocytes, and to a lesser extent its 30 kD fragment, can promote encapsulation. Phenoloxidase did not have any encapsulation-promoting activity. 相似文献
74.
Päivi Heikkilä 《Cell and tissue research》1990,259(3):421-427
Summary The present study was undertaken to define the effects of lipoprotein-derived cholesterol and endogenous, de novo synthesized cholesterol on the ultrastructure and function of undifferentiated rat adrenocortical cells [lipoprotein (HDL3 and LDL) receptor-negative, zona glomerulosa-like adrenocortical cells] in primary culture. For this purpose human plasma high density lipoprotein (HDL3) or low density lipoprotein (LDL) was added to culture medium devoid of cholesterol. Steroid secretion remained at the low basal level even after addition of lipoproteins, and the amount of intracellular lipid droplets did not increase. When mevinolin (0.96 µg/ml), an inhibitor of cholesterol synthesis, was added to the culture medium, a low secretion of corticosterone was measured both in serum-free and serum-containing media. Ultrastructurally, lipid droplets disappeared after treatment with mevinolin in both media used. At this concentration of mevinolin cell proliferation was similar to that in the controls, but at higher concentrations (4.8 or 9.6 µg/ml) proliferation was inhibited to 42% and 26% in serum-free medium, and 20% and 12% in serum-supplemented medium, respectively. This study demonstrates that cell proliferation and synthesis of corticosterone by undifferentiated rat adrenocortical cells is identical in the absence or presence of exogenous lipoprotein cholesterol. Inhibition of de novo cholesterol synthesis by mevinolin over a period of 7 days does not inhibit corticosterone secretion or proliferation of cells but decreases the amount of intracellular lipid droplets, thus suggesting utilization of intracellular cholesterol esters. However, higher concentrations of mevinolin inhibit proliferation of cells both in serum-free and serum-containing media. 相似文献
75.
Peter Bräunig 《Cell and tissue research》1990,260(1):95-108
Summary The nervus corporis cardiaci III (NCC III) of the locust Locust migratoria was investigated with intracellular and extracellular cobalt staining techniques in order to elucidate the morphology of neurons within the suboesophageal ganglion, which send axons into this nerve. Six neurons have many features in common with the dorsal, unpaired, median (DUM) neurons of thoracic and abdominal ganglia. Three other cells have cell bodies contralateral to their axons (contralateral neuron 1–3; CN 1–3). Two of these neurons (CN2 and CN3) appear to degenerate after imaginal ecdysis. CN3 innervates pharyngeal dilator muscles via its anterior axon in the NCC III, and a neck muscle via an additional posterior axon within the intersegmental nerve between the suboesophageal and prothoracic ganglia. A large cell with a ventral posterior cell body is located close to the sagittal plane of the ganglion (ventral, posterior, median neuron; VPMN). Staining of the NCC III towards the periphery reveals that the branching pattern of this nerve is extremely variable. It innervates the retrocerebral glandular complex, the antennal heart and pharyngeal dilator muscles, and has a connection to the frontal ganglion.Abbreviations
AH
antennal heart
-
AN
antennal nerves
-
AO
aorta
-
AV
antennal vessel
-
CA
corpus allatum
-
CC
corpus cardiacum
-
CN1, CN2, CN3
contralateral neuron 1–3
-
DIT
dorsal intermediate tract
-
DMT
dorsal median tract
-
DUM
dorsal, unpaired, median
-
FC
frontal connective
-
FG
frontal ganglion
-
HG
hypocerebral ganglion
-
LDT
lateral dorsal tract
-
LMN, LSN
labral motor and sensory nerves
-
LN+FC
common root of labral nerves and frontal connective
-
LO
lateral ocellus
-
MDT
median dorsal tract
-
MDVR
ventral root of mandibular nerve
-
MVT
median ventral tract
-
NCA I, II
nervus corporis allati I, II
-
NCC I, II, III
nervus corporis cardiaci I, III
-
NR
nervus recurrens
-
NTD
nervus tegumentarius dorsalis
-
N8
nerve 8 of SOG
-
OE
oesophagus
-
OEN
oesophageal nerve
-
PH
pharynx
-
SOG
suboesophageal ganglion
-
T
tentorium
-
TVN
tritocerebral ventral nerve
-
VLT
ventral lateral tract
-
VIT
ventral intermediate tract
-
VMT
ventral median tract
-
VPMN
ventral, posterior, median neuron
-
1–7
peripheral nerves of the SOG
-
36, 37, 40–45
pharyngeal dilator muscles 相似文献
76.
Marion Laig Michael Pape Joachim Hundrieser Gebhard Flatz Torpong Sanguansermsri B. M. Das Ranjan Deka Puangrat Yongvanit Naret Mularlee 《Human genetics》1990,84(2):188-190
Summary The distribution of the hemoglobin Constant Spring (Hb CS) gene in eight populations in Southeast Asia (including Assam) was determined using oligonucleotide hybridization. Hb CS was absent in two Assamese populations with a high prevalence of Hb E. The Hb CS gene frequency was 0.033 in northern Thailand and near 0.01 in central Thailand and Cambodia. High frequencies, between 0.05 and 0.06, were observed in northeastern Thailand. The present data and a similar study in Laotians suggest that the Lao-speaking populations of the Mekong River basin in northeastern Thailand and Laos have the highest frequencies of the Hb CS gene in Southeast Asia. 相似文献
77.
Günter Kämper Hans-Ulrich Kleindienst 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1990,167(2):193-200
1. | Filiform hairs of various lengths on the cerci of adult crickets vibrate in a sound field. These movements were measured with a photodetector for sound frequencies from 10 Hz to 200 Hz in the species Acheta domestica, Gryllus bimaculatus and Phaeophilacris spectrum. |
2. | With low air-particle velocities, the hair shafts were deflected sinusoidally from their resting position, without bending or secondary oscillations (Figs. 2 A, 3 A). At higher velocities (from ca. 80 mm/s peak velocity, depending on the properties of the individual hairs), the shaft struck the cuticular rim of the socket in which the base of the hair is seated (Fig. 2B). This contact was made at an average angular displacement from the resting position of 5.16°±1.0°. |
3. | The best frequencies of the hairs were found to be between 40 Hz and 100 Hz (Fig. 5A). The slope of the amplitude curve for constant peak air-particle velocity at frequencies below the best frequencies was between 0 and 6 dB/octave. Long hairs had smaller slope values than short hairs (Fig. 5C). |
4. | At its best frequency the ratio of maximal tip displacement of a hair to the displacement of the air particles in the sound field was between 0.2 and 2. Only a small number of hairs (2 out of 36) showed tip displacements exceeding twice the air-particle displacement. The values of maximal angular displacement were not correlated to hair length (Fig. 5 B). |
5. | The angular displacement of the hairs was phase shifted with respect to the air-particle velocity by 0° to +45° (phase lead) at sound frequencies around 10 Hz and by -45° to -120° (phase lag) at 200 Hz (Figs. 3C, 4B). At a particular frequency long hairs tended to have larger phase lags than shorter hairs (Fig. 5D). |
78.
Dr. Kauko K. Mäkinen Eva Söderling Salam A. Syed Pirkko-Liisa Mäkinen 《Current microbiology》1990,20(1):5-11
The cell extracts of two human oral strains (FN2 and FN3) ofFusobacterium nucleatum displayed exceptionally high-glutamylpeptidase activity as determined withN--l-glutamyl-2-naphthylamine as substrate. This activity was so dominant that the hydrolysis of otherN-aminoacyl-2-naphthylamines progressed at a rate <10% of the former. Two major enzymes (I and II) were partially purified from FN2. I had a molecular weight of 115,000 and did not hydrolyze-glutamylcysteinylglycine (glutathione). II had a molecular weight of 70,000 and rapidly liberated only glutamic acid from glutathione. Strain FN3 contained several enzymes hydrolyzing-glu-2NA. Direct anion exchange chromatography of FN3 cell extracts separated one enzyme that liberated both glutamic acid and glycine from glutathione, one that was inactive against glutathione (but hydrolyzed-glu-2NA), and one that liberated only glutamic acid. Although-glu-2NA was a good synthetic substrate, glutathione was hydrolyzed at least 500 times faster by an enzyme present in both strains. These results indicate that the presence of-glutamylpeptidase activity is very characteristic of theseF. nucleatum strains. 相似文献
79.
The effects of , , and receptor-agonists on forskolin stimulated cyclic adenosine-3, 5-monophosphate (cAMP) formation were examined in astroglial enriched primary cultures from the cerebral cortex of newborn rats. Intracellular cAMP accumulation was quantified by radioimmunoassay. Morphine was used as a -receptor agonist, D-Ala-D-Leu-Enkephalin (DADLE) as a -receptor agonist and dynorphine 1–13 (Dyn) as a -receptor agonist. Basal cAMP levels were unaffected by either the opiate agonists or the antagonists used. In the presence of the cAMP stimulator forskolin, morphine had no significant effect on the cytoplasmic cAMP levels. DADLE caused a dose related inhibition of the forskolin stimulated cAMP accumulation. The effects of this receptor stimulation was blocked with the selective antagonist ICI 174.864. In the presence of Dyn, the forskolin stimulated cAMP accumulation was inhibited in a dose related manner. This receptor stimulation was blocked with the selective antagonist MR 2266. Co-administration of DADLE and Dyn resulted in a non additive inhibition of the forskolin stimulated accumulation of cAMP. These findings indicate that astroglial enriched cultures from the cerebral cortex of rats express and -receptors co-localized ont he same population of cells, and that these receptors are inhibitory coupled to adenylate cyclase. 相似文献
80.
The enzyme acetyl-CoA: 17-O-deacetylvindoline 17-O-acetyltransferase which terminates vindoline biosynthesis has been isolated from Catharanthus roseus leaves, further characterized and purified to homogeneity by three step column chromatography and subsequent preparative isoelectric focusing. Kinetic properties concerning the enzyme reaction are discussed. Five multiple forms of the acetyl-transferase could be observed, each consisting of two subunits. This enzyme is now the best characterized of the enzymes involved in vindoline biosynthesis.Abbreviations DTE
dithiothreitol
- EDTA
ethylenediamine-tetraacetic acid
- HEPES
N-(2-hydroxyethyl)-piperazine-N-2-ethanesulfonic acid
- IEF
isoelectric focusing
- KPi
potassium phosphate
- Mr
rel.molecular mass
- PEG
polyethylene glycol
- SDS-PAGE
sodium dodecylsulfate polyacrylamide gel electrophoresis
- Tris
2-amino-2-(hydroxymethyl)-1,3-propandiol 相似文献