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Biomedical subjects

E Grzesiuk

Publications and source records attributed to E Grzesiuk.

25 records · Page 2Linked to original sources

Recombination of DNAs in Xenopus oocytes based on short homologous overlaps.

Linear molecules of pBR322 and closely related plasmid DNAs were injected into Xenopus oocyte nuclei. Such molecules were degraded unless their ends were recombined. Non-homologous ends were joined rarely, if at all, but measurable recombination was supported by homologous sequences of less than 10 base pairs (bp). The efficiency of recombination increased as the length and degree of homology improved, in the range of about 8-20 bp. The homologous sequences had to be very close to the original molecular ends (within about 20 bp); internal homologies, even when they included better matches, were never used. These observations are best accommodated by a model of recombination which envisions exonucleolytic resection to expose homologous sequences, followed by annealing of single-stranded tails, tidying up and sealing of the new joint. Some of the recombined plasmids had novel tetracycline resistance genes; their properties give some insight into the function of the tet gene product.

Animals↗

Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.

When DNA molecules are injected into Xenopus oocyte nuclei, they can recombine with each other. With bacteriophage lambda DNAs, it was shown that this recombination is stimulated greatly by introduction of double-strand breaks into the substrates and is dependent on homologous overlaps in the recombination interval. With plasmid DNAs it was shown that little or no recombination occurs between circular molecules but both intra- and intermolecular events take place very efficiently with linear molecules. As with the lambda substrates, homology was required to support recombination; no simple joining of ends was observed. Blockage of DNA ends with nonhomologous sequences interfered with recombination, indicating that ends are used directly to initiate homologous interactions. These observations are combined to evaluate possible models of recombination in the oocytes. Because each oocyte is capable of recombining nanogram quantities of linear DNA, this system offers exceptional opportunities for detailed molecular analysis of the recombination process in a higher organism.

Animals↗

Replication and expression of fragments of phage PM2 cloned in Escherichia coli K-12.

DNA from PM2 phage was cloned, as HindIII fragments and inserted into the pBR322 vector in E. coli cells. It was shown, that replication of recombined plasmids starts from the pBR322 origin. Transcription of recombinant plasmids in E. coli cells, as well as translation in minicells was demonstrated and attributed to pBR322 and/or PM2 DNA sequences.

Bacteriophages↗

[Are the cytostatic drugs and their metabolites, excreted by patients during chemotherapy, hazardous for the family environment?].

In recent years the problem concerning adverse effects of excreted drugs and their active metabolites on other people and the environment has been found to be an important aspect of clinical pharmacology and environmental health. It is commonly known that antineoplastic drugs are hazardous to medical personnel who handle them, but cytostatics administered during ambulatory care are always transferred via the patient's organism to his home and hypothetically generate potential risk to other family members. The patient s home is a sub-set of the family environment where genetically determined predisposition to neoplastic disease could additionally appear. This could intensify the effect of exogenous factors responsible for carcinogenesis (e.g. cytostatics), and increase late consequences (e.g. mutations). To assess the risk the authors elaborated an algorithm for hazard identification and assessment of consequences of the hazard. Preliminary results indicated passage a/significant amounts of excreted cytostatics and their metabolites to the natural, hospital and also family environment. After excretion, mutagenic properties of these drugs were still observed. Chemical inactivation of the excreta of patients on chemotherapy completely eliminates or strongly reduces their mutagenicity. At the present time it is difficult to assess the total risk level of cytostalic drugs and their metabolites excreted by patients on chemotherapy, for their home environment. Risk identification based on dose-response and exposure assessment, will allow to determine the health, social and economic hazards and to establish standards of procedures with regard to the risk factor.

Antineoplastic Agents↗