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Biomedical subjects

S Povey

Publications and source records attributed to S Povey.

At least 145 records · Page 8Linked to original sources

Cloning, expression and chromosomal localization of a member of the human cytochrome P450IIC gene sub-family.

We have isolated and sequenced a cDNA clone (pB8) that codes for a novel member of the cytochrome P450IIC sub-family of man. Analysis, by Southern blot hybridization, of DNA isolated from a panel of nine independent human-rodent somatic cell hybrids demonstrated that the corresponding gene (CYP2C) is located on human chromosome 10. Northern blot hybridization of RNA isolated from human livers revealed a 10-fold inter-individual variation in the expression of the gene.

Base Sequence↗

Isolation of a human cytochrome P-450 reductase cDNA clone and localization of the corresponding gene to chromosome 7q11.2.

We have isolated and sequenced cDNA clones that code for rat and human NADPH-dependent cytochrome P-450 reductase. The cDNA coding for the human protein was used to analyse, by Southern blot hybridization, DNA isolated from a panel of 8 independent human-rodent somatic cell hybrids. The results indicate that cytochrome P-450 reductase is encoded by a single gene (POR) located on human chromosome 7(pter-q22). Analysis of human metaphase chromosomes by hybridization in situ confirmed the results and refined the localization to 7q11.2. Northern blot hybridization revealed that in human liver the expression of the gene varies by less than 3-fold between different individuals.

Animals↗

Biochemical and genetic analysis of the OKa blood group antigen.

The monoclonal antibody TRA-1-85 recognizes a cell surface antigen which is expressed by all human cell types tested, including red blood cells (RBCs), but not by mouse cells. All the human RBCs tested were TRA-1-85 positive except those with the rare phenotype Ok(a-). Oka is a blood group antigen of very high frequency and only three unrelated Ok(a-) people are known. The red cells of all three propositi were negative with the TRA-1-85 antibody. To confirm the relationship between the TRA-1-85 antibody and anti-Oka, the immune antibody found in the serum of Ok(a-) individuals, Western blot analysis was used: the TRA-1-85 antibody and anti-Oka gave identical but complex patterns of reactivity in Western blot analysis of human cell lysates or membranes. This suggests that the anti-Oka and TRA-1-85 antibodies recognize the same cell-surface determinant and implies that Oka is not restricted in its expression to the surface of RBCs but is expressed on white blood cells (WBCs) of Ok(a+) individuals and all human cell lines tested to date. WBCs from one of the Ok(a-) propositi were tested and found to be negative with the TRA-1-85 antibody. Finally, the species specificity of the TRA-1-85 antibody has been exploited by the use of somatic cell hybrids and DNA transfection techniques to examine the genetic control of the Oka antigen defined by the TRA-1-85 antibody. We report that the determinant is controlled by a single gene OK present on human chromosome 19.

Animals↗

Characterization of a continuous cell line in culture established from a Krukenberg tumour of the ovary arising from a primary gastric adenocarcinoma.

A unique cell line has been established in culture from a biopsy taken from a Krukenberg tumour of the ovary arising from a primary gastric adenocarcinoma. This continuous cell line, designated KS-1, displayed epithelial-type morphological characteristics and approx. 1% of the cells had eccentric nuclei featuring the signet ring-like cells observed in the original tumour: these proved positive for neutral and acidic mucopolysaccharides. KS-1 cells have been maintained in suspension culture and had a population doubling time in logarithmic growth of approx. 28 h. They readily formed colonies in soft agar or agarose with efficiencies of 10-15%, but failed to produce tumours on inoculation into nude mice. KS-1 cells had a modal chromosome number of 60-65, with a range of 48-77%. Ten to 30 double minutes were present in most cells and several clonal marker chromosomes were identified. The effects of a 24 h in vitro exposure of KS-1 cells to a range of concentrations of 5-fluorouracil, adriamycin, mitomycin C or cisplatin have been quantitated by clonogenic assay and these values have been compared with those reported using a range of other human tumour cell lines, under comparable experimental conditions.

Adenocarcinoma↗

Genetically homozygous choriocarcinoma following pregnancy with hydatidiform mole.

Genetic studies have been made in two cases of primary choriocarcinoma from patients in whom the antecedent pregnancy was a hydatidiform mole. Restriction fragment length polymorphisms of the DNA from the tumour, the patient and her partner were examined and in both cases the tumours were shown to be androgenetic in origin, having only paternal polymorphisms. While one tumour was shown to be heterozygous, two different paternal alleles being demonstrated with some probes, the other tumour was shown to be homozygous for all informative polymorphisms examined. Thus choriocarcinoma can follow complete hydatidiform mole which may be either heterozygous or homozygous.

Adult↗

The assignment of the human gene coding for complement C5 to chromosome 9q22-9q33.

The presence or absence of the human gene for the fifth component of complement (C5) was analysed in 19 human-rodent hybrid cell lines by hybridization to a radiolabelled probe derived from a human C5 cDNA clone. The segregation of C5 in these hybrids suggested that the gene is localized on chromosome 9, in the region 9q21-9qter. In situ hybridization refined the assignment of C5 to chromosome 9q22-33.

Animals↗

The major phenobarbital-inducible cytochrome P-450 gene subfamily (P450IIB) mapped to the long arm of human chromosome 19.

We have recently isolated a cloned cDNA that codes for a human orthologue of the major phenobarbital-inducible cytochrome P450IIB subfamily of rodents. The cloned human cDNA was used to analyse, by Southern blot hybridization, DNA extracted from a panel of 9 independent human-rodent somatic cell hybrids. The results indicate that all members of the P450IIB gene subfamily of man are located on chromosome 19. Evidence from hybrids containing different regions of human chromosome 19 localizes this cytochrome P-450 gene subfamily further to the long arm of chromosome 19 in the region cen-q13.3. We propose the designation CYP2B for this locus.

Animals↗

The human alpha-1-antitrypsin-related sequence gene: isolation and investigation of its expression.

A genomic sequence approximately 5 kb in length designated the alpha-1-antitrypsin-related gene (ATR), showing considerable homology to the human alpha-1-antitrypsin (AAT) gene, was identified 10 kb downstream of the authentic AAT gene. The AAT and ATR genes were introduced separately into L-cells by transfection in order to establish a method for distinguishing between expression of the two genes. RNA probes from the cloned ATR region were then used in a ribonuclease protection assay against RNA from a range of human adult and fetal tissues. No evidence of expression of ATR was found, indicating that this region is probably a pseudogene.

Animals↗

Cloning and chromosomal mapping of human cytochrome b5 reductase (DIA1).

We have isolated a cDNA clone that codes for human cytochrome b5 reductase. The cDNA was used to analyse, by Southern-blot hybridization, DNA isolated from a panel of 11 independent human-rodent somatic cell hybrids. The results indicate that cytochrome b5 reductase is encoded by a single gene located on human chromosome 22.

Amino Acid Sequence↗

Cloning of the human alpha 1 antichymotrypsin gene and genetic analysis of the gene in relation to alpha 1 antitrypsin deficiency.

Deficiency of alpha 1 antitrypsin (Pi) is clinically heterogeneous and the unpredictability of the clinical manifestation in a person of phenotype PiZ, which may vary from severe childhood liver disease to normal health, is a problem in genetic counselling. This problem may increase as couples at risk who have not had an affected child are identified in screening programmes. One possibility is that genetic variation of other protease inhibitors may influence the prognosis. With this in mind we report the isolation of the human gene for alpha 1 antichymotrypsin (AACT) on a series of cosmid clones, with restriction mapping of about 70 kb around the gene. A probe pACE3.4 derived from the 5' end of the gene defines sequences which have been assigned to chromosome 14 using somatic cell hybrids and has been used to show a common TaqI polymorphism with allele frequencies of AACT6 = 0.7 and AACT3 = 0.3 in Europeans. pACE3.4 is closely linked to alpha 1 antitrypsin (maximum lod score in males +2.29 at theta = 0; in females Z = +6.11 at theta = 0.032). Analysis of Pi-AACT haplotypes in 31 families ascertained through PiZ or PiSZ subjects did not show any linkage disequilibrium. The distribution of AACT6 and AACT3 alleles in 16 unrelated PiZ patients presenting with childhood liver disease and five unrelated PiZ patients with adult chest disease did not differ significantly from each other. These results suggest that if genetic variation at the AACT locus does influence the outcome of alpha 1 antitrypsin deficiency, such variation is not in linkage disequilibrium with the AACT polymorphism reported here.

Cloning, Molecular↗