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Biomedical subjects

M Tully

Publications and source records attributed to M Tully.

31 records · Page 2Linked to original sources
PubMed1985

Transformation of Rhodosporidium toruloides.

Rhodosporidium toruloides protoplasts could be transformed, in the presence of polyethylene glycol (PEG), at frequencies of approx. 1 X 10(3) transformants/micrograms of DNA. The plasmid used, pHG2, which contains the phenylalanine ammonia-lyase (PAL)-coding gene (PAL) of R. toruloides, could replicate as an unstable plasmid in the yeast, or could integrate at the PAL locus to give stable transformants. Plasmids that function in R. toruloides were constructed using either the PAL gene or LEU2 gene of Saccharomyces cerevisiae as dominant selectable markers. R. toruloides transformed with pHG8, which contains both genes, coinherited the two markers. It is also shown that the 2mu replicon of S. cerevisiae does not function in R. toruloides; neither is the PAL gene expressed in S. cerevisiae.

Agaricales↗
PubMed1985

Molecular cloning of the phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli K-12.

A genomic library of Rhodosporidium toruloides DNA was constructed in bacteriophage lambda 1059. Recombinant phage containing phenylalanine ammonia lyase (PAL) gene sequences were identified by using 32P-labeled cDNA to partially purified PAL mRNA. The PAL gene was subcloned on an 8.5-kilobase PstI DNA restriction fragment into pUC8 to generate the recombinant plasmid pHG2. A restriction map of the PAL gene, together with its flanking regions, was constructed. Northern hybridization analysis of R. toruloides RNA with a restriction fragment encoding part of the PAL gene indicates that PAL mRNA is 2.5 kilobases in length. A single-stranded DNA hybridization probe was constructed and used to quantitate PAL mRNA levels in R. toruloides grown under different physiological conditions. PAL mRNA levels paralleled changes in functional PAL mRNA and antigen. These data are consistent with control of PAL expression being at the level of transcription.

Ammonia-Lyases↗
PubMed1983

Control of synthesis of functional mRNA coding for phenylalanine ammonia-lyase from Rhodosporidium toruloides.

The regulation of functional mRNA coding for phenylalanine ammonia-lyase (PAL) from Rhodosporidium toruloides was investigated. Polyadenylic acid [poly(A)]-containing RNA was an efficient template for in vitro translation in rabbit reticulocyte lysate. Non-poly(A)-containing RNA did not stimulate in vitro protein synthesis. Several lines of experimental evidence indicate that mRNA from R. toruloides directs PAL synthesis in reticulocyte lysate: (i) the major radioactive product in immunoprecipitates when lysates, incubated with yeast poly(A)-containing RNA, were reacted with PAL-antiserum had the same molecular weight as native PAL (75,000); (ii) this major radioactive product competes with authentic PAL for binding to PAL-antiserum; and (iii) partial proteolytic peptide maps of the in vitro translation product were very similar to those of native PAL. The levels of functional mRNA coding for PAL, when R. toruloides was grown in different physiological conditions, were determined by quantitation of PAL synthesized in vitro when RNA was added to reticulocyte lysate. Functional PAL mRNA was six times higher in yeast grown on phenylalanine compared with glucose-phenylalanine minimal medium. No functional PAL mRNA was detected in yeast grown on glucose-ammonia minimal medium in the presence or absence of phenylalanine. These observed changes in functional PAL mRNA were similar to levels of PAL catalytic and antigenic activity. The kinetics of functional PAL mRNA synthesis and degradation were studied. Maximum levels of functional PAL mRNA were observed within 60 min of transfer to PAL-inducing growth conditions. Poly(A)-containing RNA and functional PAL mRNA were rapidly degraded when cells were transferred from phenylalanine to glucose-ammonia minimal medium, with half-lives of 25 and 10 min, respectively. Thus, it is suggested that the alterations in the amount of PAL in cells of R. toruloides grown in different physiological conditions primarily result from alteration in the amount of functional mRNA coding for the enzyme.

Ammonia-Lyases↗
PubMed1983

Child's play.

Explore the source record for details and available documents.

Child Day Care Centers↗
PubMed1982

Synthesis and degradation of phenylalanine ammonia-lyase of Rhodosporidium toruloides.

The regulation of the enzyme phenylalanine ammonia-lyase (PAL), which is of potential use in oral treatment of phenylketonuria, was investigated. Antiserum against PAL was prepared and was shown to be monospecific for the enzyme by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The native enzyme and two inactive mutant forms of the enzyme were purified to homogeneity by immunoaffinity chromatography, using anti-PAL immunoglobulin G-Sepharose 4B. Both mutant enzymes contained intact prosthetic groups. The formation of PAL catalytic activity after phenylalanine was added to yeast cultures was paralleled by the appearance of enzyme antigen. During induction, uptake of [3H]leucine into the enzyme was higher than uptake into total protein. Our results are consistent with de novo synthesis of an enzyme induced by phenylalanine, rather than activation of a proenzyme. The half-lives of PAL and total protein were similar in both exponential and stationary phase cultures. No metabolite tested affected the rate of enzyme degradation. Glucose repressed enzyme synthesis, whereas ammonia reduced phenylalanine uptake and pool size and so may repress enzyme synthesis through inducer exclusion. The synthesis of enzyme antigen by a mutant unable to metabolize phenylalanine indicated that this amino acid is the physiological inducer of the enzyme.

Ammonia↗
PubMed1980

Chronic pulmonary infection with Salmonella typhimurium.

The first report of a case of chronic sputum carriage of salmonella is presented. The patient was a 51-year-old male with pulmonary cavitation and mycetoma, who developed pulmonary infection with Salmonella typhimurium after rectal surgery, and subsequently has carried the salmonella in his sputum for 18 months to date.

Ampicillin↗
PubMed1980

Bacteriostasis of Escherichia coli by milk. VI. The in-vitro bacteriostatic property of Gambian mothers' breast milk in relation to the in-vivo protection of their infants against diarrhoeal disease.

A one-year field-study has been carried out in a diarrhoea-endemic area in West Africa to determine the relationship between the bacteriostatic activity of fresh human milk for Escherichia coli in vitro and freedom from diarrhoea of the infant recipients of the milk. The specific contribution of E. coli gastroenteritis to gastrointestinal diseases of infants in general is not known, nor is its particular role in the Gambian infants studied. During the study period, however, both enteropathogenic and toxigenic strains of E. coli were isolated. The incidence of diarrhoea in Gambian infants of seven age-groups from 2 days to 12 months was not significantly correlated with the bacteriostatic activity of milk. This was due rather to absence of diarrhoea in babies fed on low-activity milk than illness in those receiving highly bacteriostatic milk. Indeed, very active milk appeared to protect recipients almost completely, including seven babies of over 3 months of age, five of them during the rainy season, when the risk was high. Babies receiving lower-activity milk experienced more diarrhoea. In a situation where diarrhoeal disease is multifactorial, field evaluation of the protective action by one antibacterial property of milk is difficult. A better understanding of in vivo protection is important, and the factors which have to be taken into account are discussed.

Diarrhea↗
PubMed1977

Fine-structure mapping and complementation analysis of the Escherichia coli cysB gene.

Sixty-two point mutations were isolated in Escherichia coli by means of transduction with mutagenized phage P1. Twenty-two deletions extending into cysB but able to recombine with at least some of the point mutations were isolated on a transmissible E. coli plasmid. Mapping of the point mutations against the deletions divided the former into 16 deletion groups. Nine merodiploids were constructed in which the chromosome carried one of the three point mutations most distal to the trp operon and in which a plasmid carried one of the three point mutations most proximal to the trp operon. All of these showed a Cys-phenotype. It follows that mutations at the two extreme ends of the region belong to the same complementation group.

Chromosome Mapping↗
PubMedDate not supplied

A memory clinic for the assessment of mild dementia.

The aims and experience of work in a specialist memory clinic' are described with reference to the first 200 patients attending the service. 93.5% of attenders were found to suffer from dementia. 67.9% of these fulfilled criteria for probable/possible Alzheimer's disease or other primary degenerative dementias and 8.6% fulfilled criteria for vascular dementia. In the remaining cases the aetiology of the dementia was considered to be multifactorial. The memory clinic proved to be an important resource for demented patients, their carers, and for research projects.

Aged↗