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J W Dale

Publications and source records attributed to J W Dale.

At least 55 records · Page 3Linked to original sources
PubMed1989

Oxacillin-hydrolysing beta-lactamases. A comparative analysis at nucleotide and amino acid sequence levels.

We have extended the sequence of the OXA-2 beta-lactamase which together with S1 mapping has enabled us to identify the promoter site for this gene. This lies in a region that is found upstream from a variety of resistance genes on different plasmids; each gene appears to have been inserted at the same specific site and to be expressed from the same promoter. The ancestral plasmid thus appears to function as a natural expression vector. The sequence of the recombination site at the 5' end of the OXA-2 gene shows a marked similarity with the attP sequence of lambda. DNA-probe analysis confirmed that the OXA-2 and OXA-3 beta-lactamases are related, and indicated no similarity with other beta-lactamase genes. However, a comparison of amino acid sequences demonstrates that the OXA-2, OXA-1 and PSE-2 beta-lactamases show some similarities to the typical class A enzymes, especially in the central helical domain of the latter, which is largely responsible for forming the active site of the enzyme. The three oxacillinases also show marked amino acid sequence similarity with the product of a regulatory gene, blaR1, required for beta-lactamase induction in Bacillus licheniformis.

Amino Acid Sequence↗
PubMed1989

Polymorphic repetitive DNA sequences in Mycobacterium tuberculosis detected with a gene probe from a Mycobacterium fortuitum plasmid.

Clinical isolates of Mycobacterium tuberculosis were shown by Southern blotting to contain DNA sequences hybridizing to a probe derived from a Mycobacterium fortuitum plasmid. Two such M. tuberculosis DNA fragments, isolated from a gene library, were used as probes to show restriction fragment length polymorphism in M. tuberculosis strains by detecting a repetitive sequence apparently located at different points on the chromosome. This could indicate the presence of a transposable element in M. tuberculosis which is partly homologous to a region of the M. fortuitum plasmid. The probes described can be used to fingerprint M. tuberculosis isolates, and in addition are capable of distinguishing M. tuberculosis from Mycobacterium bovis and BCG.

Blotting, Southern↗
PubMed1989

Transformation of Mycobacterium smegmatis with Escherichia coli plasmids carrying a selectable resistance marker.

One limiting factor in studies of tuberculosis and leprosy is the difficulty of genetic analysis and manipulation of mycobacteria. Two approaches were adopted for the construction of vectors, based on different Escherichia coli plasmids and using Mycobacterium smegmatis as the host. In both cases we found that the original E. coli plasmid is capable of being replicated in M. smegmatis, yielding chloramphenicol-resistant colonies. One such plasmid has been recovered from a M. smegmatis transformant and used to re-transform both M. smegmatis and E. coli to chloramphenicol resistance. This plasmid is indistinguishable from the original plasmid by restriction analysis, and can be used as a shuttle vector for the genetic manipulation of mycobacterial species.

Blotting, Southern↗
PubMed1989

Transformation of Mycobacterium smegmatis with E. coli plasmids.

One limiting factor in the studies of tuberculosis and leprosy is the lack of a versatile system for genetic analysis and manipulation of mycobacteria. One strategy used in constructing a plasmid vector for transforming Mycobacterium smegmatis was to insert fragments of a mycobacterial plasmid into an Escherichia coli plasmid. We found that the parental E. coli plasmid is capable of self-replication in M. smegmatis yielding chloramphenicol-resistant colonies. Plasmids from different passages of one M. smegmatis transformant were recovered and characterised by restriction digest analysis. The plasmid from the earlier passage was found to be indistinguishable from the original plasmid by restriction analysis. Plasmids from later preparations, however, were found to have undergone modifications in the M. smegmatis host resulting in an apparent increase in transformation efficiency for M. smegmatis. These plasmids can be used as a shuttle vector for the genetic manipulation of mycobacterial species.

DNA, Bacterial↗
PubMed1988

Differential splicing in the 3' non-coding region of rat cytochrome P-452 (P450 IVA1) mRNA.

The gene coding for the clofibrate-induced cytochrome P-452 has been isolated from a cDNA library and a full-length cDNA clone sequenced. The sequence was identical with that reported by Hardwick et al. [(1987) J. Biol. Chem. 262, 801-810] apart from the presence of an additional 75 nucleotides inserted adjacent to the 3'-end of the coding region. Northern blot analysis indicates that this is probably a consequence of differential splicing, which may be relevant to the tissue-specific regulation of this gene.

Amino Acid Sequence↗
PubMed1986

Cloning and expression of Mycobacterium bovis BCG DNA in "Streptomyces lividans".

The ability of "Streptomyces lividans" to use the expression signals of genes from Mycobacterium bovis BCG was tested in vivo by using gene fusions. Random DNA fragments from M. bovis BCG were inserted into promoter-probe plasmids in Escherichia coli and in "S. lividans." Comparison with promoter activity detected with random DNA fragments from the respective hosts suggested that "S. lividans" efficiently utilizes a high proportion of mycobacterial promoters, whereas a smaller fraction are expressed, and expressed more weakly, in E. coli. M. bovis BCG DNA fragments were also inserted into the specially constructed translational fusion vector (pIJ688) in "S. lividans." pIJ688 contains the kanamycin phosphotransferase gene (neo) from transposon Tn5, truncated at its amino terminus, as the indicator. The results suggested that "S. lividans" uses M. bovis BCG translational signals almost as efficiently as its own signals. Moreover, several hybrid proteins with an M. bovis BCG-derived amino terminus seemed to be reasonably stable in "S. lividans." These experiments indicate that "S. lividans" may be a suitable host for the expression of Mycobacterium leprae and Mycobacterium tuberculosis genes from their own signals. This is a precondition for the expression of entire biosynthetic pathways, which could be valuable in the production of diagnostic and therapeutic agents. The vectors may also have wider applications for the analysis of gene expression in Streptomyces.

Cloning, Molecular↗
PubMed1985

Sequence of the OXA2 beta-lactamase: comparison with other penicillin-reactive enzymes.

The nucleotide sequence of the unusual plasmid-mediated OXA2 beta-lactamase is presented, and compared with other beta-lactamases. The OXA2 enzyme has similar features at the presumed active site, but no other significant regions of homology with other penicillin-reactive enzymes. The active site homology may therefore represent convergent evolution of otherwise dissimilar genes.

Amino Acids↗
PubMed1985

Escherichia coli thymine auxotrophs suppressible by carbon dioxide.

Two independently-isolated thymine-requiring mutant strains of Escherichia coli were found to possess an unusual phenotype: in the presence of CO2, growth was independent of thymine. The two strains showed different profiles of temperature sensitivity. Glycine, serine and methionine were unable to relieve the thymine-dependence. Both strains were susceptible to trimethoprim under conditions where they were thymine-independent. The results are consistent with the occurrence of partial defects in thymidylate synthase.

Carbon Dioxide↗
PubMed1984

Improved purification and characterization of the OXA-2 beta-lactamase.

An improved and scaled-up procedure has been developed for purifying the OXA-2 plasmid-mediated beta-lactamase. This has enabled us to improve the characterization of this enzyme, including a revised determination of its amino acid composition and the sequence of the N-terminal region of the protein.

Amino Acids↗
PubMed1984

Antenatal care in high-risk pregnancies.

A retrospective analysis of the antenatal records of women who gave birth to live babies was conducted in the three Districts of the East Sussex Health Area in 1977. A high-risk group was identified and the antenatal care for them was compared with that given to mothers in general. Comparison was also made between Districts. Women in the high-risk group were similar to those in the general group with respect to the number of antenatal visits. Overall this was also found with respect to when the last visit was made, but there were differences among those with short or long gestations. Pregnant women at high risk were more likely to be supervised by the hospital team exclusively during the antenatal period than those in the general group. Maternal characteristics which might have been used to identify high-risk pregnancies were often not recorded. High-risk mothers more frequently had infants with Apgar scores of less than 5 at one minute and birth weights of 2500 g or less than did women in the general group. However, it would appear that District policy on health care varied significantly since various aspects of care were found to be more dependent on the District concerned than on the risk group involved.

Adult↗
PubMed1982

The interaction of anthraquinone dyes with the plasmid-mediated OXA-2 beta-lactamase.

Although beta-lactamases do not require any nucleotide co-substrates, the OXA-2 type is inhibited competitively by Cibacron Blue 3GA, and by other anthraquinone dyes, including some simpler compounds with no side chain. The enzyme causes a red shift in the spectrum of Cibacron Blue. The beta-lactamase can be adsorbed in Blue Sepharose and specifically eluted by benzylpenicillin. These results indicate that the binding of anthraquinone dyes is a specific effect similar to that seen with many nucleotide-binding enzymes.

Anthracenes↗
PubMed1979

The effect of a plasmid on growth and survival of E. coli.

We studied the growth characteristics of a pair of Escherichia coli strains, isogenic apart from the possession of a nonconjugative plasmid. There was no difference between the two strains when they were grown separately. In mixed culture, a second slow phase of growth that normally occurred following the end of rapid exponential growth, was absent from the plasmid-carrying strain. This resulted in a considerable decrease in the proportion of the cells that carried the plasmid after overnight incubation. The effect of different conditions of growth is reported. The plasmid-carrying strain survived extended incubation (150 days at 37 degrees C) as well as did the plasmid free strain separately. In a mixture, the proportion of plasmid-carrying cells declined rapidly, and none was detected after 100 days.

Cell Division↗
PubMed1978

High and low level tetracycline resistance in Shigella sonnei.

The results presented in this paper confirm the existence of two types of tetracycline resistance in Shigella sonnei. One group of strains had a high level of resistance to tetracycline and oxytetracyline, with a variable level of minocycline resistance. The second group had a lower level of tetracycline resistance and were sensitive to minocycline. After conjugation with E. coli K12 the selected E. coli transconjugants had the same levels of resistance as the parent Sh. sonnei strain, with one exception. Sh. sonnei 87 was resistant to a high level of tetracycline, but was able to transfer only low level resistance. It is suggested that Sh. sonnei 87 carriers two plasmids: pSU1, a conjugative plasmid conferring a low level of tetracycline resistance, and pSU2, a non-conjugative plasmid which confers a high level of resistance to tetracycline.

Microbial Sensitivity Tests↗
PubMed1977

Semiprecious alloys for cast restorations: a preliminary report.

A previous clinical evaluation indicated that semiprecious metals had a definite place in the construction of dowels and cores in endodontically treated teeth. 2 The results from the present in vitro study suggest that those alloys do have a place in the fabrication of single castings as type III and IV gold alloy substitutes. Although when compared with a type III gold alloy the behavior of the semiprecious alloys tested was less like that of a noble alloy, the semiprecious alloys appeared less active than the base-metal alloy. This suggests that the semiprecious alloys should be acceptably resistant to corrosion in the oral environment.

Crowns↗
PubMed1977

A clinical evaluation of semiprecious alloys for dowels and cores.

The semiprecious metals investigated and hard dental gold alloys may be manipulated in a similar manner. The semiprecious metals produced accurate and well-fitted dowels needing minimal adjustment. Being harder than type III gold alloy, they would be ideal for casting dowels and cores. Before considering these semiprecious alloys as final restorations, however, they must be further examined for resistance to tarnish and corrosion and their marginal adaptability and burnishability.

Crowns↗