PubMed Health⌕ Search

Biomedical subjects

A Orosz

Publications and source records attributed to A Orosz.

At least 19 recordsLinked to original sources
PubMed1991

Analysis of the complex transcription termination region of the Escherichia coli rrnB gene.

The complex terminator region of the Escherichia coli rrnB gene was analyzed by subcloning the terminators T1 and T2 and the inverted repeats IR1 and IR2 individually, or in various combinations, in a normal or inverted orientation into a terminator probe vector. The in vivo terminating efficiency was assayed by measuring the galactokinase activity encoded by the downstream galK gene. Termination efficiencies of all fragments were compared in two constructs, differing in the presence or absence of readthrough translation over the investigated terminator signal. The following main conclusions were drawn. (a) T1 and T2 are both efficient terminators in isolated forms. (b) IR1 and IR2 have some terminating effect (much lower than the proper terminators), especially in the inverted orientation. Their presence modifies the effect of the proper terminators in a quite unpredictable way, especially if these regions are translated. (c) The terminators are not symmetrical; in the inverted orientation T1 is practically inactive and T2 termination is reduced. (d) Translation radically decreases the efficiency of the terminators. (e) Several sequences in the rrnB gene, upstream of the terminator region (one in the 16S RNA and one in the 5S RNA coding region), are very efficient in vivo terminators in the inverted orientation.

Base Sequence↗
PubMed1990

A family of expression vectors based on the rrnB P2 promoter of Escherichia coli.

We describe here the construction of a family of expression vectors, based on the P2 promoter of the Escherichia coli rrnB gene by removing regulatory sequences downstream of the Pribnow-box and replacing them with the lac operator. These vectors allow cloning of foreign genes in such a way that their products are synthesized either in the form of fusion proteins of different length, or without fusion partners, with or without the original translational initiation signals. One of the vectors contains a synthetic oligothreonine-coding sequence that helps to stabilize the product of the cloned gene. These vectors allow high-level regulated expression of foreign genes, even if their products are relatively short peptides.

Amino Acid Sequence↗
PubMed1990

New approaches to increase the expression and stability of cloned foreign genes in Escherichia coli.

A family of expression plasmid vectors were constructed by fusing the strong P2 promoter of the rrnB gene of Escherichia coli (coding for ribosomal RNA) to the lac operator, thereby eliminating regulatory sequences from the rrnB gene and placing the expression under lac repressor control. This promoter proved to be stronger in vivo than the well-known consensus tac promoter, and its strength could be further increased by converting the sequence to consensus. The stability of the recombinant proteins could be increased by fusion to various lengths of the N-terminal end of beta-galactosidase, or by inserting a synthetic oligonucleotide, coding for heptathreonine. A new method was developed for the stabilization of recombinant plasmids without antibiotic selection, based on the presence of an essential gene on the plasmid and its absence from the chromosome. The application of this method is illustrated by the example of a plasmid expressing human proinsulin.

Biotechnology↗
PubMed1989

Combined effects of fasting and vinblastine treatment on serum insulin level, the size of autophagic-lysosomal compartment, protein content and lysosomal enzyme activities of liver and exocrine pancreatic cells of the mouse.

1. The volume fraction of autophagic vacuoles in liver parenchymal and exocrine pancreatic cells was smallest and the serum insulin level highest in the 24 hr prestarved mouse immediately after 3 hr feeding period. 2. The size of the autophagic vacuole and lysosome (dense body) compartments increased in both types of cells during 2-72 hr fasting parallel with decreasing serum insulin levels. 3. The protein content of the cells decreased and the DNA-based activity of acid phosphatase showed little change throughout fasting. The activity of cathepsin D increased during days 2 and 3 of food deprivation. 4. Vinblastine (50 mg/kg body wt) applied for the last 2 hr of different periods (2, 12, 24, 48 and 72 hr) of fasting decreased serum insulin level and increased the fractional cytoplasmic volume of autophagic vacuoles and dense bodies. This increase was smaller when the drug was applied shortly after feeding and much larger after prolonged fasting. The increase was more pronounced in the pancreatic than in the liver cells. 5. Our data show that the effect of vinblastine on the size of the autophagic-lysosomal compartment depends on the feeding status of the animals.

Acid Phosphatase↗
PubMed1988

BcefI, a new type IIS restriction endonuclease.

A new Type IIS restriction endonuclease was identified, partially purified and characterized from a Bacillus cereus subsp. fluorescens strain. The enzyme recognizes the nonpalindromic sequence ACGGC and cleaves at a distance from it. The cleavage appears to occur with a +/- 1 basepair uncertainty. Thus the cleavage and recognition site is as shown below: ACGGC(N)11-13 TGCCG(N)12-14.

Bacillus cereus↗
PubMed1988

Alteration of insulin-binding receptors in non-insulin dependent diabetes of the young.

The behaviour of insulin binding receptors is rather unelucidated in non-insulin-dependent diabetes mellitus of the young. Authors in continuing their previous work studied the behaviour of insulin binding receptors of erythrocytes and monocytes in 9 MODY patients. They observed that specific insulin binding of circulating blood cells was significantly decreased in all cases as compared to the controls despite of a good state of metabolism (in the case of erythrocytes 4.63 +/- 1.1% vs. 6.03 +/- 1.7%, p less than 0.05, in the case of monocytes 2.3 +/- 1.2% vs. 3.6 +/- 1.4%, p less than 0.05). The lower value of insulin binding resulted from the decrease of receptor concentrations (in the case of erythrocytes 2.36 +/- 0.78 pmol/l vs. 3.81 +/- 1.14 pmol/l, p less than 0.05).

Adult↗
PubMed1983

Separation of synaptic junctional complexes from thin layers of rabbit cerebral cortex.

Synaptic junctional fractions were separated from rabbit brain by procedures based on combining the methods of Cotman and Taylor [4], Orosz et al. [16, 17] and Lisman et al. [13]. Thin layers of cerebral cortices were homogenized to obtain a crude mitochondrial-synaptosomal fraction. The sedimentation rates of mitochondria and mitochondria containing synaptosomes were increased by raising the density of mitochondria with an insoluble dense formazan deposit inside mitochondria after iodo-nitrotetrazolium treatment. The synaptic plasma membrane fraction isolated by this method contained no mitochondrial contamination. After Triton X-100 treatment the insoluble residues of the detergent were centrifuged through discontinuous sucrose gradients. A great enrichment of morphologically identifiable intact synaptic junctions was observed in some of the obtained interface layers.

Animals↗
PubMed1975

Synaptosomal fractions of cat cerebral cortex after hypo-osmotic and detergent treatment.

Different synaptosomal fractions were prepared by subcellular fractionation from cat cerebral cortex. The seperated nerve ending fractions differed in both density and osmotic sensitivity. Synaptosomal ghosts were obtained by hypo-osmotic treatment. In the synaptosomal ghost fractions the synaptic areas seemed to be intact morphologically. The morphology of the "synaptic triad" (pre- and postsynaptic membranes and the synaptic cleft) was not affected even by treatment with Triton X-100 non-ionic detergent (0.6%). The solubilizing effect of different concentrations of the detergent was checked in the supernatants and in the insoluble residues. The homogeneity of various subcellular fractions and the effect of the osmotic and detergent treatments are discussed.

Animals↗