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1.
Guanylate cyclase from crude homogenates of vegetative has been characterized. It has a pH optimum of 8.0, temperature optimum of 25°C and requires 1 mM dithiothreitol for optimal activity. It strongly prefers Mn++ to Mg++ as divalent cation, requires Mn++ in excess of GTP for detectable activity, and is inhibited by high Mn++ concentrations. It has an apparent Km for GTP of approximately 517 μM at 1 mM excess Mn++.The specific activity of guanylate cyclase in vegetative homogenates is 50–80 pmoles cGMP formed/min/mg protein. Most of the vegetative activity is found in the supernatant of a 100,000 x g spin (S100). The enzyme is relatively unstable. It loses 40% of its activity after 3 hours storage on ice. Enzyme activity was measured from cells that had been shaken in phosphate buffer for various times. It was found that the specific activity changed little for at least 8 hours. Cyclic AMP at 10?4 M did not affect the guanylate cyclase activity from crude homogenates of vegetative or 6 hour phosphate-shaken cells. 相似文献
2.
J.A. Pérez-González A. Jiménez 《Biochemical and biophysical research communications》1984,125(3):895-901
The paromomycin producing organism is resistant to this antibiotic and contains a phosphotransferase which inactivates paromomycin. The gene encoding this enzyme has been inserted in the vector pIJ702 and then cloned in , selecting for paromomycin-resistance. Three plasmids have been isolated and one of them, pMJ1, contains a 2.2 kb insert with a single HindIII restriction site. Insertion of foreign DNA in this site blocks the expression of the phosphotransferase enzyme indicating that it is within the cloned gene. These findings provide a new dominant selective marker for cloning vectors with the versatility of insertional inactivation. 相似文献
3.
Robert C. Richmond John D. Zimbrick 《Biochemical and biophysical research communications》1975,64(1):391-398
Cells of C thy?321 are examined for thymine residue release from DNA following gamma-irradiation from 5 to 15 krad. Experimental conditions are designed to inhibit enzyme activity that might promote base residue release. Enzyme action is restricted in order to assess the physicochemical action of radiation on cellular DNA, and to this end irradiation is done under O2, N2, and N2O saturating conditions. Both thymine and thymidine release from bacterial DNA are detected and quantitated, and three oxygen effects are noted in comparing yields of these products. No difference in effect is observed between N2 and N2O gassing conditions, suggesting that the hydroxyl radical has little effect on thymine or thymidine release from irradiated DNA . 相似文献
4.
Ronald M. Hamelik Mead M. McCabe 《Biochemical and biophysical research communications》1982,106(3):875-880
An inhibitor of , endodextranase was detected in proteins prepared from batch cultures of , strains representing serotypes through . Affinity chromatography of strain 6715-49 proteins, which apparently were free of endodextranase activity, yielded an active endodextranase and, in a separate peak, the endodextranase inhibitor. The presence of the inhibitor in culture fluids accounts for the absence of endodextranase activity in batch-grown cultures of , known to produce this enzyme. 相似文献
5.
Evelyn A. Devine Mary C. Moran Peter J. Jederlinic Anthony J. Mazaitis Henry J. Vogel 《Biochemical and biophysical research communications》1975,67(4):1589-1593
The transducing phage λd, carrying a portion of the chromosome including , is derived from the heat-inducible, lysis-defective strain λy199, which has the and deletions. Cleavage of λy199 DNA by RI endonuclease, followed by agarose slab gel electrophoresis, results in bands corresponding to the known C, D, E, and F segments of λ, and a segment A′ (A plus B minus minus , the cleavage site between A and B being eliminated). Cleavage of λd DNA by RI yields the expected D, E, and F segments of λ and four other segments, termed 14-1 through 14-4, whose length is 17.5, 6.2, 3.0, and 2.0 kilobases, respectively, as determined by electron microscopy and corroborated by electrophoretic mobility. Heteroduplex analysis shows that the cluster is on the 14-1 segment. 相似文献
6.
Yuji Kamiya Akira Sakurai Nobutaka Takahashi 《Biochemical and biophysical research communications》1980,94(3):855-860
Rhodotorucine which induces mating tube formation of cells in is metabolized rapidly by cells. By use of labeled rhodotorucine , the degradation was found to be proteolytic. Two peptide fragments Tyr-Pro-Glu-Ile-Ser-Trp-Thr-Arg and Asn-Gly-Cys(S-farnesyl) were identified as the metabolites. Proteolysis of the pheromone mainly occurred on the cell surface. Culture filtrate of cells at log phase did not metabolize rhodotorucine . 相似文献
7.
Yolk free blastoderms of chick embryo were incubated 3 or 22 hours with labeled pregnenolone, progesterone, 17-hydroxyprogesterone, dehydro-epiandrosterone, androstenedione, testosterone and estradiol-17β. Metabolites and unconverted substrates were found both in the incubation medium and in the cells. Enzymes responsible for identified conversions were: 17α-hydroxylase, 17-20-desmolase, Δ53β- and 3α-hydroxysteroid dehydrogenase, 17β-hydroxysteroid dehydrogenase and 5α- and 5β-reductase. The results suggest that the steroid metabolizing enzyme activities found may reflect a more general ability of early embryonic cells. 相似文献
8.
OKY-1581 is an effective inhibitor of thromboxane synthesis and . The generation of thromboxane B2 (TxB2), prostaglandin E (PGE) and prostaglandin F (PGF) was measured following clotting and during platelet aggregation induced by collagen. The presence of OKY 1581 either or caused a reduction in TxB2 generation during clotting and platelet aggregation with a concomitant increase in PGE and PGF. The effect could be observed two hours after oral or subcutaneous administration of 5 to 100 mg per rabbit and lasted for 24 to 48 hours. The reduction in TxB2 was not accompanied by an inhibition of clotting or platelet aggregation. OKY-1581 appears to be a suitable agent for studying the role of TxB2 in atherosclerosis. 相似文献
9.
An enzyme which catalyzes the reduction of bisulfite to sulfide and thiosulfate was purified from extracts of the sulfate-reducing bacterium, . Trithionate was not a product of this reaction nor was it or thiosulfate reduced by the enzyme. High substrate concentrations inhibited sulfide but not thiosulfate formation. The enzyme was named bisulfite reductase II to distinguish it from bisulfite reductase which reduces bisulfite to trithionate. 相似文献
10.
W. Wackernagel 《Biochemical and biophysical research communications》1973,51(2):306-311
Genetic transformation of for various chromosomal markers was accomplished by (i) using recipient cells that lack the DNase but were recombination proficient due to or mutations and (ii) treating the recipient cells with CaCl2 at a concentration that facilitates transfection by λ DNA. Cotransformation of three markers () was found to depend on the molecular weight of the transforming DNA. 相似文献
11.
Josie M. Nichols G.A.M. Cross 《Biochemical and biophysical research communications》1976,71(3):796-802
Kinetoplast DNA from the order Kinetoplastidae (trypanosomatids) exists as large associations (molecular weight 4 × 1010), made up of about 104 small, probably circular, molecules, commonly known as ‘minicircles’. These minicircles were originally thought to be identical in base composition, suggesting that the coding capacity of kinetoplast DNA is very restricted. However, linear molecules have also been observed in preparations of kinetoplast DNA, which, if they contain unique sequences, could represent additional genetic information. This linear DNA has been assumed to be derived from the kinetoplast, but the possibility of it being nuclear contamination has not been definitely ruled out. Work presented in this paper demonstrates that nuclear DNA contamination may indeed be present in kinetoplast DNA prepared by a commonly used method. 相似文献
12.
This case report addresses the occurrence of Brucellosis and its effect on the cattle in developing countries. Three Zebu bulls () are presented and the clinical and pathologic signs are described. Conception rates declined following an abortion storm in one herd and without prior abortions in another herd. Semen collected by electro-ejaculation was found to be azoospermic or with very few spermatozoa. was isolated from seminal vesicles, testes and epididymides. Organs affected and showing microscopic lesions were testes, epididymides and seminal vesicles. The latter were not consistently affected. None of the bulls showed impairment of libido or breeding capacity. 相似文献
13.
G. Sheir-Neiss R.V. Nardi M.A. Gealt N.R. Morris 《Biochemical and biophysical research communications》1976,69(2):285-290
[35S] labeled extracts of the fungus were copolymerized with purified porcine brain tubulin. The [35S] protein which copurified with porcine microtubules was found to be similar to [3H] chick tubulin when the two were coelectrophoresed on several polyacrylamide gel electrophoresis systems. These results strongly suggest the presence in of a tubulin-like protein. 相似文献
14.
James B. Howard Thomas Lorsbach Lawrence Que 《Biochemical and biophysical research communications》1976,70(2):582-588
In 80% dimethyl sulfoxide/H2O, ferredoxin FeS clusters can be extruded with benzene thiol. The extruded clusters have an absorption spectra maximum at 458 nm which is characteristic of 4Fe4S centers. The amino terminal sequence of the ferredoxin has 7 of the 8 Cys residues at residue numbers 8, 11, 16, 20, 24, 39 and 42. Except for Cys 24, all of these residues can be correlated to homologous Cys residues in other bacterial ferredoxins. Although two thirds of the first 45 residues are identical to or conservative replacements for the first 43 residues of other bacterial ferredoxins, the insertion of Cys-24 indicates a major change in the environment of one of the two 4Fe4S clusters. 相似文献
15.
Uterine stage embryos collected from the hamster (8-cell) and cow (morula, early blastocyst) were monitored for development (embryo culture) and (embryo transfer) following premature removal of the zona pellucida.Removal of the zona pellucida did not significantly affect development to the blastocyst stage of (1) 8-cell hamster embryos (zonae removed by a combined enzymic-mechanical procedure), (2) bovine morulae (zonae removed by mechanical means only) (3) early bovine blastocysts (zonae removed by the enzymic-mechanical technique).Zona-free hamster embryos formed significantly fewer viable fetuses than did zona-intact embryos. The lower incidence of fetal development observed following transfer of zona-free 8-cell hamster embryos may have resulted in part from the formation of chimeras by fusion of these embryos . Such fusion was observed to occur between zona-free embryos placed in close proximity. The proportion of pregnancies resulting from transfer of bovine blastocysts cultured from zona-free morulae was similar to that of zona-intact embryos.In this study we have demonstrated that (1) enzymic and mechanical procedures used to remove zonae pellucidae from uterine-stage hamster and bovine embryos do not adversely affect subsequent development of these embryos and and (2) zonae pellucidae are not required for normal development of these embryos. These findings have implications for microsurgery of mammalian embryos and for embryo transfer. 相似文献
16.
The specific synthesis of F mRNA directed by the F gene carried on the specialized transducing bacteriophage F, performed , is described with the use of an S180 extract from a strain carrying R?. Synthesis of F mRNA is biphasic at approximately 7 minutes. The regulation of F mRNA synthesis by the specific arginine holorepressor present in an S180 extract prepared from a strain carrying the R+ allele is described. 相似文献
17.
F. Malpartida M. Zalacaín A. Jiménez J. Davies 《Biochemical and biophysical research communications》1983,117(1):6-12
The gene encoding the phosphotransferase enzyme that modifies hygromycin B in its producing organism , has been cloned in the vector pIJ41. Two plasmids, pFM4 and pFM6, containing 2.1 and 19.6 kb inserts of DNA, respectively, which express the modifying enzyme, have been isolated. A 3.1 kb PstI restriction fragment from pFM4 was inserted in the vector pIJ350 and the resulting plasmids, pMZ11.1 and pMZ11.2, express the hygromycin B-resistance phenotype. The utility of this dominant marker for cloning experiments is discussed in the text. 相似文献
18.
In an attempt to isolate and to study the electron transport system of , we have isolated and purified a membrane-bound cytochrome . The cytochrome , purified as a detergent (Triton X-100) and hemoprotein complex, contained 1.6 nmoles heme per mg of protein. Cold-temperature spectrum showed that no other cytochrome was associated with the purified preparation, and electrophoresis revealed that only one type of hemoprotein was obtained. The purified cytochrome reacted with both carbon monoxide and cyanide readily. Only in the reduced form did it combine with carbon monoxide, whereas the oxidized form reacted with cyanide. An “oxygenated” form of the cytochrome was demonstrated to be spectrally distinguishable from both the oxidized and the reduced forms. 相似文献
19.
The synthesis and characterization of - and -3,4-bis(4-hydroxyphenyl)-2-hexene (- and -pseudo-DES) and of -3,4-bis(4-hydroxyphenyl)-2-hexen-1-ol (-1-hydroxypseudo-DES) are described. These compounds are useful as probes in the study of hormone action. 相似文献
20.
P. Bertrand M. Bruschi M. Denis J.P. Gayda F. Manca 《Biochemical and biophysical research communications》1982,106(3):756-760
Cytochrome c553 is a monohaemic c type cytochrome isolated from the sulfate reducing bacteria ,. Its midpoint potential value, determined by optical, EPR and polarographic studies is significantly lower than the midpoint potentials reported for other monohaemic cytochromes c (+ 10 mV instead of + 290 mV). In an attempt to study correlations between amino acid sequence, haem iron coordination and haem exposure in cytochromes c, cytochrome c553 is compared with mitochondrial and bacterial c type cytochromes. 相似文献