Penicillinase inactivates kanamycin in the presence of carbenicillin.
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Biomedical subjects
Publications and source records attributed to W Klingmüller.
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Azospirillum brasilense, A. amazonense, and A. lipoferum strains were screened for restriction endonucleases using phage lambda DNA. The extract of A. brasilense 29711 cleaved lambda DNA into specific fragments. It was concluded that this strain possesses a class II restriction endonuclease which was named AbrI. AbrI has a single recognition site on lambda DNA at position of approx. 33 500 bp. AbrI was characterized as an isoschizomer of XhoI, which cuts lambda DNA at 33 498 bp and cleaves double-stranded DNA at the sequence 5'-C TCGAG-3'. From other Azospirilla strains only A. amazonense QRZ42 extracts (AamI activity) cleaved DNA into specific fragments under certain conditions.
The nif gene group from Klebsiella can be transferred into Enterobacter cloacae by conjugation using Escherichia coli donor cells carrying the composite self-transmissible nif-plasmid pRD1. A small fraction of the hybrids obtained is stable upon prolonged passaging without selection. Their stability is due to integration of pRD1 into the chromosome. Such integration hybrids were chlorate resistant, and nitrate reductase negative, which indicated that integration preferentially occurred within one of the genes for the production or functioning of this enzyme. Chlorate resistance could, therefore, be used to select for additional nitrate reductase-negative sublines with pRD1 in their chromosome. Such sublines have been analyzed further for the presence of nif genes, other pRD1 markers, and for stability. In all except one the complete plasmid seems to have been integrated. Some tend to revert to nitrate utilisation (chlorate sensitivity).
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Examples for genetic engineering are the transfer of nuclei between cells of higher animals and the introduction of heterologous DNA into bacteria by means of plasmids. The former approach will help to establish new ways in animal breeding, the latter provides bacterial cells that produce proteins of medical importance. The moral justification of related studies in man is still open, but the possible risks of gene technology can be coped with by adhering to proper safety regulations.
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Genetic engineering has ushered in a new era in biology. Although many problems are still to be solved, there are examples that point to a possible later application for the benefit of mankind: Bacteria can be manipulated to degrade crude-oil spillages, to produce human insulin and to bind nitrogen from the air. If all the bacteria that are indigenous to agricultural soils could be made to bind nitrogen, an increase in soil fertility might well result.
Bacterial resistance factors are extrachromosomal elements, consisting of circular closed, double stranded DNA. They may contain genes which make the host bacterium resistant to certain antibiotics. Such resistance factors have become important vectors for selective replication of specific segments of all kinds of DNA. Suitable DNA segments are obtained by treating DNA with restriction endonucleases of defined cutting properties. They can then be linked to the respective vector. It was thus possible to anneal DNA segments originating from the same, but also from different bacterial resistance factors. They gave biologically functional units. The annealed products can be propagated in bacteria. The relevant experiments are described. Their potential dangers for man, and efforts to agree on suitable safety regulations are discussed.
The effect of edeine and the mutation edr-2 to edeine resistance on genetic recombination in Neurospora crassa was investigated. For this purpose crosses between pairs of edeine sensitive and edeine resistant strains respectively were set up without or in the presence of the drug (0-750 microg/ml). The genetic markers ylo-1, ad-1, pan-2 (B3 and B5) and tryp-2, all on linkage group VI, were used for scoring recombinants. These were ad+, tryp+ (intergenic recombination) and pan+ (interallelic recombination). Frequencies of about 6-7% for intergenic and of about 0.4% for interallelic recombination were found in crosses between eds strains and ed(r) strains respectively, if edeine was absent. However, crosses in the presence of edeine gave higher frequencies of both intergenic and interallelic recombination (about 12% intergenic and 1% interallelic with 180 to 200 microg ed/ml). The pan+ prototrophs (interallelic recombinants) obtained in the different crosses were tested for distribution of outside markers. The data thus obtained revealed that under the effect of both the mutation to edeine resistance and edeine itself the relative number of noncrossover (gene conversion) recombinants decreases in favour of crossover recombinants, and the relative number of double crossover recombinants (events outside the pan locus) decreases in favour of single crossover recombinants. It is concluded that a) edeine and the mutation ed(r)-2 to edeine resistance affect recombination via related pathways, and b) noncrossover and crossover recombinants are caused by different molecular mechanisms, in agreement with the work of other authors.
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