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1.
N. Gilboa-Garber V. Zakut L. Mizrahi 《Biochimica et Biophysica Acta (BBA)/General Subjects》1973,297(1):120-124
Production of cholinesterase by a pyocyanin-producing strain of Pseudomonas aeruginosa, isolated from a patient and grown in the presence of acetylcholine as the main source of carbon, was described. The enzyme activity was detected in suspensions of intact bacteria and in their subcellular preparations. Like the acetylcholinesterase of the electric eel, as opposed to that of the erythrocytes, this bacterial enzyme was inhibited by specific antiserum produced against it in rabbits. The production of the enzyme was found to be sensitive to catabolite repression and to require external cyclic AMP, but not 5′-AMP for the elimination of this repression. Cyclic AMP alone, without the inducer, did not stimulate the production of the enzyme. 相似文献
2.
Kinetic mechanism of DNA polymerase I (Klenow) 总被引:12,自引:0,他引:12
The minimal kinetic scheme for DNA polymerization catalyzed by the Klenow fragment of DNA polymerase I (KF) from Escherichia coli has been determined with short DNA oligomers of defined sequence. A key feature of this scheme is a minimal two-step sequence that interconverts the ternary KF.DNAn.dNTP and KF.DNAn+1.PPi complexes. The rate is not limited by the actual polymerization but by a separate step, possibly important in ensuring fidelity [Mizrahi, V., Henrie, R. N., Marlier, J. F., Johnson, K. A., & Benkovic, S. J. (1985) Biochemistry 24, 4010-4018]. Evidence for this sequence is supplied by the observation of biphasic kinetics in single-turnover pyrophosphorolysis experiments (the microscopic reverse of polymerization). Data analysis then provides an estimate of the internal equilibrium constant. The dissociations of DNA, dNTP, and PPi from the various binary and ternary complexes were measured by partitioning (isotope-trapping) experiments. The rate constant for DNA dissociation from KF is sequence dependent and is rate limiting during nonprocessive DNA synthesis. The combination of single-turnover (both directions) and isotope-trapping experiments provides sufficient information to permit a quantitative evaluation of the kinetic scheme for specific DNA sequences. 相似文献
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The apparent involvement of ornithine decarboxylase (ODC) and putrescine in the early stages of fruit growth in tomato (Lycopersicon esculentum Mill.) has been previously described. Further evidence presented here supports the direct involvement of ODC and putrescine in the cell division process in tomato fruits. In tomato fruits grown in vitro, in which basic growth processes are inhibited, the activity of ODC and arginine decarboxylase (ADC) and the level of free polyamines were reduced. While ODC and ADC activity was correlated with the period of cell division in the tomato fruit, the free polyamine content was correlated with the DNA content, cell size, and fruit fresh weight. The addition of exogenous putrescine, however, did not restore the basic growth processes in the fruits grown in vitro. 相似文献
6.
Investigation into the nature of a Bacillus promoter cloned into a promoter-probe plasmid 总被引:1,自引:0,他引:1
The alpha-amylase-coding gene (amy) of Bacillus amyloliquefaciens NCP1 was cloned into the Bacillus subtilis promoter probe vector pPL603b.1, using a BglII digest of chromosomal DNA. The resulting plasmid, pVC102, was shown to have a BglII site within the insert. It was determined that this was the result of the fortuitous co-cloning of 2.88-kb and 0.92-kb BglII fragments separated in NCP1 DNA by approx. 3 kb. Unexpectedly, this co-cloning was readily repeated. Subcloning showed that while the 2.88-kb amy-bearing fragment was sufficient for amylase production, it might not have been capable of promoting sufficient levels of chloramphenicol resistance under the conditions used in the cloning experiments. The promoter on the 0.92-kb BglII fragment was more efficient, although its sequence differed from the canonical promoter sequence recognised by B. subtilis RNA polymerase E.sigma 43. As other promoter-bearing fragments from NCP1 DNA operated equally efficiently when cloned into pPL603b.1, the reason for the repeated co-cloning of the 2.88-kb and 0.92-kb NCPI BglII fragments may well be due to structural parameters, whereby certain nucleotide sequences are more readily cloned than others. 相似文献
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The Uba2 and Ufd1 proteins of Saccharomyces cerevisiae interact with poly(A) polymerase and affect the polyadenylation activity of cell extracts 总被引:2,自引:0,他引:2
Poly(A) polymerase is responsible for the addition of the adenylate tail to the 3′ ends of mRNA. Using the two-hybrid system,
we have identified two proteins which interact specifically with the Saccharomyces cerevisiae poly(A) polymerase, Pap1. Uba2 is a homolog of ubiquitin-activating (E1) enzymes and Ufd1 is a protein whose function is probably
also linked to the ubiquitin-mediated protein degradation pathway. These two proteins interact with Pap1 and with each other,
but not with eight other target proteins which were tested in the two-hybrid system. The last 115 amino acids of Uba2, which
contains an 82-amino acid region not present in previously characterized E1 enzymes, is sufficient for the interaction with
Pap1. Both Uba2 and Ufd1 can be co-immunoprecipitated from extracts with Pap1, confirming in vitro the interaction identified
by two-hybrid analysis. Depletion of Uba2 from cells produces extracts which polyadenylate precursor RNA with increased efficiency
compared to extracts from nondepleted cells, while depletion of Ufd1 yields extracts which are defective in processing. These
two proteins are not components of polyadenylation factors, and instead may have a role in regulating poly(A) polymerase activity.
Received: 6 January 1997 / Accepted: 27 February 1997 相似文献
9.
The marine bacteria Beneckea harveyi and Photobacterium leiognathi were shown to bear mannose-containing binding sites for the mannosephilic lectins of Pseudomonas aeruginosa and concanavalin A (Con A). The interaction between the lectins and the marine bacteria was demonstrated by the bacteriagglutination test, by adsorption of the lectins onto the bacteria and by mannose-specific peroxidase-binding to the lectin-coated bacteria. Treatment of the bacteria with formaldehyde, phenol, ethanol or boiling them for 15 min, did not alter their ability to adsorb the lectins. The growth rate of the marine bacteria was unaffected when either the Pseudomonas lectins or Con A was added to the culture medium. 相似文献
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