Trimethoprim resistance controlled by a combination of plasmid and chromosomal genes.
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Host ranges of members of four groups of male-specific RNA coliphages were determined by plating on hosts carrying various derepressed plasmids. An RNA phage originally isolated on Pseudomonas aeruginosa failed to form plaques on any of the strains of Escherichia coli.
Transferable chloramphenicol resistance has become common in the typhoid bacillus in countries such as Mexico, India, Vietnam and Thailand. Situations such as this, and others analogous to it in many parts of the world, are the result of the long-term indiscriminate use of chloramphenicol and other antibiotics in the affected areas. They can be rectified only by more rational antibiotic usage.
There are situations in which direct observation of the attachment of antibody molecules may be the simplest method of detecting antigen-antibody reactions. Applied to the study of the sex pili determined by a number of transmissible bacterial plasmids, the method has distinguished four serotypes in the F-like class and two in the I-like class. Antibody was usually attached haphazardly to the pili; however, in a few cases a regular periodicity could be observed. When few antibody molecules were attached, they could frequently be individually resolved and in certain antibody-pilus combinations large antibody molecules, tentatively identified as IgM, could be seen to predominate.
The antibiotic resistance patterns of coliforms in faecal specimens from pigs and their human contacts were studied. The ability of the resistant coliforms to transfer their resistance in vitro to antibiotic-sensitive recipients was examined. The results showed that pigs which had received antibiotics carried more multiply-resistant, R-factor bearing coliforms than pigs which had not been given antibiotics. Human contacts of the antibiotic-treated pigs had a higher incidence of antibiotic-resistant coliforms with R-factors than human contacts of pigs which had not been given antibiotics. It is concluded that antibiotic treatment of farm animals may lead to acquisition of antibiotic resistance by gut coliforms of man.
A case of typhoid fever caused by Salmonella typhi occurred in Geneva. The patient was probably infected in Mexico City. The strain isolated from this patient corresponds with the description of the Mexican S. typhi strain, since it is a degraded Vi-strain resistant to chloramphenicol, streptomycin, sulphonamides and tetracyclines. It carried an fi(-) transferable R factor with a CSSuT resistance pattern. It can be accepted that this case forms part of the Mexican outbreak of chloramphenicol-resistant typhoid fever which has already been observed in visitors to Mexico from England and the United States.
The transfer of an R factor from donor E. coli introduced into the rumen of adult sheep to strains of the coliform microflora resident post rumen in the lower gastro-intestinal tract was found to be greatly increased when the animals were subjected to a short period of starvation (ca. 24-48 h). This also resulted in coliform organisms containing the resistance determinants of the R factor being excreted for much longer periods, sometimes for months afterwards. As no antibiotic treatment was given to the animals during these experiments, possession of the R factor should have conferred no selective advantages on the host cells and other plasmids could possibly be transferred similarly in vivo in sheep or other ruminants and perhaps also within the gut of monogastric animals.
During the winter of 1967-8 Sonne dysentery affected neighbouring North London primary schools. This was not simply due to cross-infection between the two schools, for two different, unusual strains of Sh. sonnei were distinguished. One was a novel kanamycin-resistant colicine type 7 strain, and as far as we know this was the first school outbreak due to such a strain to be documented. The other strain was kanamycin-sensitive and of colicine type 0 with a rare specific requirement for aspartic acid. There was some evidence to suggest that the kanamycin-resistant strain was more infective for adults and possibly more pathogenic than the kanamycin-sensitive.Studies on the transfer of drug resistance and of colicinogeny revealed that the factor determining colicine type 7 was carried on a transmissible plasmid, a new observation. Various drug resistances were also transmissible experimentally, and some were spontaneously unstable. Non-transferable ampicillin-resistance in the colicine type 7 strain and aspartic acid dependence in the colicine type 0 strain enabled all but one of the isolates to be classified into two distinct lines. No common ancestor was found and it was concluded that although occurring together they must have arisen from separate sources.
Forty Theileria schizont-infected lymphocyte culture isolates from Zimbabwe were characterized using a panel of antischizont monoclonal antibodies (MAbs) and 4 Theileria parva DNA probes containing cloned extrachromosomal element, Tpr repetitive, ribosomal and telomeric sequences. The Theileria isolates were assigned as T. parva or T. taurotragi on the basis of reactivities with MAbs and restriction fragment length polymorphisms (RFLPs) detected using the extra-chromosomal element probe. Cattle-derived T. parva isolates were relatively homogeneous on the basis of reactivities with MAbs and RFLPs detected using Tpr repetitive and ribosomal DNA probes. In contrast to previous results from Kenya, most of the cattle-derived isolates from Zimbabwe exhibited very similar Tpr restriction fragment patterns, although the Tpr genotypes of buffalo-derived isolates were heterogeneous. This suggests that selection for a particular Tpr genotype may be occurring in cattle. Many isolates with similar Tpr genotypes were differentiated by RFLPs detected using the telomeric DNA probe. The T. parva Boleni immunizing stock was distinguished from all other isolates by telomeric RFLPs. The T. parva Boleni Tpr repetitive DNA probe cross-hybridized with T. taurotragi DNA and detected RFLPs between different T. taurotragi isolates.
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Treatment of a 5-bromodeoxyuridine-resistant (brdUrdr) and prolactin-nonproducing (Prl-) subclone of GH cells with this drug led to amplification of the prolactin (Prl) gene and induced Prl synthesis. Withdrawal of the drug treatment reversed both of these processes. In normal rats, the increased Prl synthesis observed during late pregnancy and lactation does not seem to be mediated via amplification of the gene. Amplification of the Prl gene and induction of Prl synthesis can also be observed in the Prl-, brdUrd-sensitive (brdUrds) GH cell strain. Prl gene amplification thus does not seem to be associated with the mechanism that confers the brdUrdr phenotype to these cells. brdUrd-induced amplification of the Prl gene can be identified with the low molecular weight, extrachromosomal, supernatant DNA fraction, isolated by Hirt's method. Southern blot analysis of Hirt's supernatant DNA (undigested) from brdUrd-treated cells generated a distinct band following hybridization with [32P]pDNAPrl-insert. The size of this band is greater than 23 kb but smaller than chromosomal DNA. Growth hormone (Gh) and albumin (Alb) gene sequences can be detected in the chromosomal DNA preparation but are absent in the extrachromosomal DNA prepared from Hirt's supernatant. The levels of Gh and Alb sequences are unaffected by brdUrd treatment of these cells. Results presented here suggest that in rat pituitary glands as well as in GH cells, hormonally controlled increased Prl synthesis is not caused by gene amplification. However, the brdUrd-induced expression of the Prl gene seems to be linked to the mechanism of drug-induced amplification of the Prl gene, mediated via an extrachromosomal event.
The positions of the several sea urchin histone genes on the eukaryotic fragments of the chimeric plasmids pSp2 and pSp17 have been mapped relative to the Eco RI and Hind III restriction endonuclease sites on the plasmids. Two principal mapping methods using the electron microscope have been used: (a) the R-loop procedure and a new modification thereof to map the genes on duplex DNA; (b) the gene 32-ethidium bromide technique to visualize RNA-DNA hybrids on single strands of DNA. It is known that there are two histone genes, H3 and H2A, on pSp17. There are two Eco RI sites at the two junctions of the procaryotic segment with the eucaryotic segment on the plasmid. We show, by an electron microscope method, that for H2A, with a length of 0.52 kilobases (kb), one end of the gene is situated 0.02 to 0.03 kb from one RI site, and that there is a Hind III site within this gene at about 0.13 kb from the end phe other RI site of this plasmid. The H4 gene lies between H2B and H1. The ms the incubation temperature is raised up to a temperature just below that at which strand dissociation of the duplex DNA occurs.
A chimeric plasmid has been constructed in vitro from colicin E1 factor (Col E1), nontransmissible R-factor RSF-1010, and Drosophila melanogaster DNAs by the sequential action of Escherichia coli endonuclease RI(Eco RI) and T4 phage DNA ligase. The chimeric plasmid was assembled in two stages--first, a composite plasmid consisting of Col E1 and RSF 1010 was constructed, followed by partial digestion of the composite with Eco RI (in order to open one of the susceptible cleavage sites) and ligation with an Eco RI-digested D. melanogaster DNA preparation. The chimeric plasmid was selected and amplified in vivo by sequential transformation of E. COLI C with the ligated mixture, selection of transformants in medium containing streptomycin plus colicin E1, followed by amplification in the presence of chloramphenicol and purification of the extracted plasmid by dye-buoyant density gradient centrifugation in ethidium bromide-CsCl solution. Treatment of the chimeric plasmid with Eco RI yields three fragments with mobilities corresponding to the linear forms of the constituents--COL E1, mol wt 4.2 times 106, RSF 1010, mol wt 5.5 times 106 and D. melanogaster DNA, mol wt 4.0 times 106. The buoyant densities of the three constituents are respectively 1.706, 1.719, and 1.697 g/cm3, while the buoyant density of the composite factor is 1.712 and that of the chimeric plasmid is 1.705. Serratia marscesens endonuclease R (Sma) which introduces a single cut in Col E1, but not in RSF 1010, converts the chimeric plasmid to a single linear molecule (mol wt 13.7 times 106) and sequential digestion with both Sma and Hin III yields two distinct fragments, mol wt 3.7 and 10.0 times 10.6, respectively; this implies that the two sites are unique and occur at distinctly different positions. Sequential digestion with both Hin III and Eco RI reveals that the Hin III cut is in the D. melanogaster segment; neither Col E1 nor RSF 1010 contain sites susceptible to digestion with Hin III. In the presence of chloramphenicol, the chimeric plasmid continues toreplicate for 9 hr while bacterial chromosomal DNA replicates at a much slower rate. As in the case of the composite plasmid, continued synthesis is the presence of chloramphenicol suggests that the replicator of Col E1 is functional in the chimeric plasmid as well. Examination of the chimeric plasmid by partial denaturation mapping permits identification of its constituents, each of which presents a characteristic profile. The D. melanogaster segment reveals a wealth of detail at the molecular level pertaining to the distribution of AT-rich regions.
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