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Biomedical subjects

E P Green

Publications and source records attributed to E P Green.

11 recordsLinked to original sources
PubMed1993

Determination of the organisation of coding sequences within the iduronate sulphate sulphatase (IDS) gene.

The structure of the gene coding for iduronate sulphate sulphatase (IDS) has been determined. We have used exon to exon and vectorette PCR to identify 9 exons within the IDS gene and to characterise the surrounding intron sequences. The results of this study will be useful for the complete analysis of many mutations giving rise to Hunter syndrome. IDS is the first member of the group of lysosomal nonarylsulphatase genes for which the gene structure has been determined. It bears no relationship to the exon organisation of steroid sulphatase, despite the homology between these two proteins. This suggests that the division of the sulphatases into the two subgroups on the basis of substrate specificity is also reflected at the level of gene structure.

Base Sequence↗
PubMed1992

Detection of point mutations and a gross deletion in six Hunter syndrome patients.

We have used screening with the polymerase chain reaction and chemical mismatch detection of amplified cDNA to detect and characterize deletions and point mutations in six Hunter Syndrome patients. A high degree of mutational heterogeneity was observed. The first patient is completely deleted for the gene coding for alpha-L-iduronate sulfate sulfatase, while the second has a point mutation that creates a stop codon. The third patient shows a point mutation that creates a novel splice site that is preferentially utilized and results in partial loss of one exon in the RNA. Patients 4, 5, and 6 have point mutations resulting in single amino acid substitutions. Four of the six single-base changes observed in this study were examples of transitions of the highly mutable dinucleotide CpG to TpG. This study has demonstrated a procedure capable of detecting all types of mutation that affect the function of the IDS protein and should enable direct carrier and prenatal diagnosis for Hunter syndrome families.

Amino Acid Sequence↗
PubMed1992

IS902, an insertion element of the chronic-enteritis-causing Mycobacterium avium subsp. silvaticum.

An insertion sequence element of Mycobacterium avium subsp. silvaticum was isolated and its complete nucleotide sequence determined. IS902 is 1470 bp in size and is repeated 10-12 times per genome. An open reading frame of 1200 bp was identified, encoding a protein product of Mr 43932. This protein is highly similar to the predicted proteins of IS900 of Mycobacterium paratuberculosis, IS116 of Streptomyces clavuligerus and IS110 of Streptomyces coelicolor. IS902 lacks terminal inverted repeats and flanking direct repeats but displays insertion site specificity.

Amino Acid Sequence↗
PubMed1991

Specific detection of Mycobacterium paratuberculosis by DNA hybridisation with a fragment of the insertion element IS900.

This paper describes the evaluation of a newly developed DNA probe for Mycobacterium paratuberculosis. DNA probe PCR278 is a 278 bp fragment obtained by polymerase chain reaction (PCR) amplification of the 5'-region of IS900, an insertion element contained in the genome of M paratuberculosis. This DNA probe can specifically distinguish M paratuberculosis from a wide range of other organisms, including members of the M avium-M intracellulare complex. When used in conjunction with the PCR amplification technique DNA probe PCR278 could detect as little as 10 fg (equivalent to two genomes) starting material of M paratuberculosis genomic DNA. Use of PCR amplification assays based on IS900, for the detection of M paratuberculosis, and homologous IS elements found in disease isolates of M avium should greatly help our understanding of the role of these organisms in Crohn's disease and other chronic inflammatory disorders.

Base Sequence↗
PubMed1989

Sequence and characteristics of IS900, an insertion element identified in a human Crohn's disease isolate of Mycobacterium paratuberculosis.

The complete sequence of an insertion element IS900 in Mycobacterium paratuberculosis is reported. This is the first characterised example of a mycobacterial insertion element. IS900 consists of 1451bp of which 66% is G + C. It lacks terminal inverted and direct repeats, characteristic of Escherichia coli insertion elements but shows a degree of target sequence specificity. A single open reading frame (ORF 1197) coding for 399 amino acids is predicted. This amino acid sequence, and to a lesser extent the nucleotide sequence, show significant homologies to IS110, an insertion element of Streptomyces coelicolor A3(2). It is proposed that IS900, IS110, and similar insertion elements recently identified in disease isolates of Mycobacterium avium are members of a phylogenetically related family. IS900 will provide highly specific markers for the precise identification of Mycobacterium paratuberculosis, useful in defining its relationship to animal and human diseases.

Amino Acid Sequence↗
PubMed1989

DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients.

Strains of the Mycobacterium avium intracellulare complex (MAIC) have become important colonisers of patients with acquired immunodeficiency syndrome (AIDS). Restriction fragment length polymorphisms were used to study the DNA from 88 MAIC isolates, including 51 derived from 47 AIDS patients. MAIC isolates from 33 of 45 AIDS patients were identical at the molecular level and distinct from the mycobacteria isolated from the stools of healthy subjects. The study also showed that serotyping correlates poorly with the genetic identity of these organisms. Mycobacterium paratuberculosis, which has been implicated in Crohn's disease, was not identified in any of the cultures studied.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Inhibition of antral gastrin cells by peptide histidine isoleucine.

The present studies were directed to determine whether peptide histidine isoleucine (PHI), like its structural analogues secretin and gastric inhibitory peptide, inhibits antral gastrin. In separate experiments, the effects of PHI on medium gastrin concentrations, the incorporation of [35S]methionine into newly synthesized gastrin, and steady-state gastrin mRNA were determined. The inclusion of PHI in the incubation medium decreased medium gastrin levels at all concentrations examined, an effect that was not altered by the addition of 10(-6) M tetrodotoxin to the medium. PHI also inhibited the incorporation of [35S]methionine into newly synthesized gastrin in a concentration-dependent manner. Steady-state levels of gastrin mRNA were determined by dot-blot hybridization, using a 32P-labeled gastrin cRNA probe. PHI inhibited gastrin mRNA levels in a concentration-dependent manner; in contrast, no effect on the levels of actin and ubiquitin mRNA could be detected, indicating specificity of PHI on gastrin mRNA. The results of these studies indicate that PHI may exert a physiological inhibitory effect on antral gastrin cells and that this inhibition may occur at several steps along the biosynthetic pathway.

Animals↗
PubMed1989

Insertion elements in mycobacteria.

We have isolated and characterised a repetitive element from the genome of Mycobacterium paratuberculosis. This repetitive element shows many features characteristic of a bacterial insertion element.

Cloning, Molecular↗
PubMedDate not supplied

Living therapy.

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