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Biomedical subjects

A J Clark

Publications and source records attributed to A J Clark.

At least 289 records · Page 16Linked to original sources
PubMed1983

Genetic analysis of transposon-induced mutations of the Rac prophage in Escherichia coli K-12 which affect expression and function of recE.

Fourteen mitomycin-resistant revertants of a recB21 recC22 strain were isolated after Tn5 mutagenesis. Eight of the mutations (type I) were essentially inseparable from aphA+ (Kanr) of Tn5; six (type II) were not. We hypothesize that the former are Tn5 and that the latter are IS50 insertions. Because of their phenotypic similarity to sbcA and sbcB mutations, which also suppress recB21 recC22, we have called them sbc mutations. sbc-lll::Tn5 was cotransducible with nirR and has thereby been located at position 29.8 on the Escherichia coli map in the vicinity of the Rac prophage and sbcA mutations. A recB21 recC22 sbc-lll::Tn5 strain was subjected to Tn10 mutagenesis, and a mitomycin- and UV-sensitive mutant was isolated. tet+ of Tn10 was 85% cotransducible with aphA+ of Tn5, locating these two transposons 0.1 map unit apart. A three-point cross located the Tn10 mutation at position 29.7. We hypothesize that the Tn10 insertion is located in recE and that the Tn5 and IS50 insertions activate expression of this gene. sbc-lll::Tn5 was found to be cis acting and dominant to its wild-type allele as were two sbcA mutations (sbcA1 and sbcA6). Five other type I and type II insertion mutations were dominant to their wild-type alleles. We hypothesize that the sbc insertion and sbcA mutations affect transcription regulation of recE and discuss the possibility that they do so differently.

Alleles↗
PubMed1983

Restriction nuclease and enzymatic analysis of transposon-induced mutations of the Rac prophage which affect expression and function of recE in Escherichia coli K-12.

Fourteen Tn5-generated mutations of the Rac prophage, called sbc because they suppress recB21 recC22, were found to fall into two distinct types: type I mutations, which were insertions of Tn5, and type II mutations, which were insertions of IS50. Both orientations of Tn5 and IS50 were represented among the mutants and were arbitrarily labeled A and B. All 14 of the Tn5 and IS50 insertions occurred in the same location (+/- 100 base pairs) approximately 5.6 kilobases from one of the hybrid attachment sites. Eleven of the mutants contained essentially the same amount of exonuclease VIII, the product of recE. The possibility that a promoter for recE was created by the insertion of Tn5 and IS50 was considered. Two IS50 mutants in which such a promoter could not have been created showed three to four times as much exonuclease VIII, and another showed one-half as much as the majority. The possibility was considered that a promoter internal to IS50 is responsible for this heterogeneity. Restriction alleviation was measured in all 14 mutants. An insertion of the transposon Tn10 which reduces expression of exonuclease VIII (recE101::Tn10) was located within the Rac prophage at a position 2.35 kilobases from the left hybrid attachment site. Location and orientation of the Rac prophage on the Escherichia coli genetic map are discussed in light of these results.

Coliphages↗
PubMed1982

Biosynthetic human insulin in the treatment of diabetes. A double-blind crossover trial in established diabetic patients.

94 diabetic patients established on treatment with porcine (n = 47) or bovine (n = 47) insulin took part in a double-blind crossover trial, in which 6-week periods of treatment with the appropriate animal insulin were compared with periods of treatment with biosynthetic human insulin (BHI). 6 patients withdrew during the trial, in 3 cases because of hypoglycaemia while taking BHI. In bovine-insulin-treated patients, the mean glucose level (mean of seven capillary-blood samples over 1 day), the modified M index, and total daily insulin requirement were the same on BHI and bovine-insulin treatment. For porcine-insulin-treated patients, mean glucose level and the modified M index were slightly higher on BHI than on porcine-insulin treatment (9.7 vs 9.0 mmol/l and 79.6 vs 65.0, respectively), despite an average increase of 2.3 units/day of BHI after 6 weeks of such treatment. Hypoglycaemic episodes were not significantly more or less frequent on BHI in either group of patients. In both groups fasting blood glucose was higher during BHI treatment than during animal-insulin treatment (14.2 vs 12.8 mmol/l [bovine group]; 12.1 vs 9.6 mmol/l [porcine group]). In bovine-insulin-treated patients blood glucose before the evening insulin injection was higher on BHI than on bovine insulin (11.6 vs 10.0 mmol/l). BHI appears to be a safe alternative to porcine or bovine insulin. Differences in the pharmacokinetics of BHI may account for the observed differences in blood-glucose responses.

Adult↗
PubMed1982

Two main groups of mouse major urinary protein genes, both largely located on chromosome 4.

Fourteen different major urinary protein (MUP) genomic clones from BALB/c mice were isolated. By restriction site mapping, six of these form two sets of three overlapping clones. By the criterion of cross-hybridization, the 10 different genes fall into two groups of four (Group 1) and three (Group 2) genes, while three genes fall into neither group. Southern blot analysis of genomic DNA with Group 1 and Group 2 plasmid subclones shows that the haploid mouse (BALB/c) genome contains approximately 15 Group 1 genes, 12 Group 2 genes and at least seven MUP genes that belong to neither group. An analysis of mouse-Chinese hamster hybrid cell lines shows that most, if not all, Group 1 and Group 2 genes are located on mouse chromosome 4.

Animals↗
PubMed1982

Effects of a recA operator mutation on mutant phenotypes conferred by lexA and recF mutations.

Derepression of recA by an operator mutation (recAo281) produces effects opposite to those obtained from its derepression following DNA damage. Inducible reactivation of lambda vir and S13 phages is decreased and inducible UV mutagenesis of a phi X174 amber mutant is lessened in a recAo281 strain compared to a recAo+ strain. The decreases could not be accounted for by increases in constitutive levels of these processes. Consistent with these results the UV resistance of a recAo281 strain is less than that of a recAo+ strain. This may indicate that too much recA protein immediately after irradiation interferes with derepression of the lexA regulon or functioning of its products. Effects of increasing the recAo+ and recA+ copy number on a Co1E1 plasmid are compared with the effects of recAo281. recAo281 partially suppresses UV sensitivity due to lexA102 and lexA3 in E. coli K-12. This increase in resistance is not correlated with an increase in constitutive or inducible reactivation of UV-irradiated lambda vir or S13. This is consistent with the previous suggestion that the UV resistance stems from a decrease in DNA degradation allowing an increase in DNA repair. lexA3 blocks UV mutagenesis of phi X174 as measured by reversion of amber mutations and this was not suppressed by recAo281. recF143 blocks UV mutagenesis of phi X174. recAo281 suppresses neither this effect nor the decrease in bacterial UV resistance caused by recF143.

Bacteriophage phi X 174↗
PubMed1982

Variation between mouse major urinary protein genes isolated from a single inbred line.

We describe ten Charon 4A genomic DNA clones from BALB/c mice which include at least seven different major urinary protein (MUP) genes. We have established the orientation of all seven sequences, and have placed six of them in precise register by means of restriction site maps and Southern blot hybridization with cloned cDNA sequences. Four of the seven genomic sequences (family I sequences) form hybrids with six independent cDNA clones that have a high thermal stability and hybridize more strongly with mRNA from three inbred mouse lines. Hybrids between the remaining three genomic sequences and the cDNA clones have a lower thermal stability and hybridize less strongly with mRNA from the three inbred lines. Homologies between different cloned sequences extend over as much as 15 kb. No clone contains parts of two MUP genes, and no homology has been detected between the 3' flanking region of one MUP gene and the 5' flanking region of another.

Animals↗
PubMed1981

The measurement of membrane potential during photosynthesis and during respiration in intact cells of Rhodopseudomonas capsulata by both electrochromism and by permeant ion redistribution.

1. The membrane potential in intact cells of Rhodopseudomonas capsulata during photosynthesis and during dark respiration has been measured by two independent methods. 2. The light-induced and O2-induced shifts in the carotenoid absorption spectrum were measured in the intact cells. The shift was calibrated with K+-diffusion potentials in chromatophores derived from those cells. The light-induced and O2-induced membrane potentials were -290 mV and -230 mV respectively. 3. The energized uptake of butyltriphenylphosphonium ions was measured in the same batch of cells. The light-induced and O2-induced membrane potentials calculated from the Nernst equation were -160 mV and -120 mV respectively. 4. It is concluded that the two kinds of probe measure the electric potentials across different domains of the cytoplasmic membrane, but it is difficult to reconcile the existence of such domains with simple electrical analogues of the membrane and aqueous phases.

Bacterial Chromatophores↗
PubMed1981

Physical mapping of the srl recA region of Escherichia coli: analysis of Tn10 generated insertions and deletions.

A restriction endonuclease map for the enzymes EcoRI, BamHI, SalI, and PstI covering 23.5 kilobase pairs (kb) of the srl recA region of Escherichia coli was constructed. An insertion of the transposon Tn10 in the negative regulatory gene srlR was shown to be located 5.8 kb away from the promoter proximal end of the recA gene. The extent of several Tn10 generated deletions, originating from the srlR301::Tn10 insertion, were analyzed by physical mapping. Three mutations that had removed the Tn10 encoded tetracycline resistance gene, del(srl-recA)302, del(srl-recA)304, and del(srl-recA)303, were found to be deleted for 40%, 45%, and 50% of the recA structural gene, respectively. A deletion, del(srl-recA)306, that had not affected the structure of the Tn10 in srlR301 was shown to have removed the entire recA structural gene.

Chromosome Deletion↗
PubMed1981

Premedication with meperidine and atropine does not prolong recovery to street fitness after out-patient surgery.

The effect of premedication with meperidine and atropine on recovery to street fitness after out-patient surgery under general anaesthesia was assessed. The subjects of this study were 100 female patients undergoing therapeutic abortion. Two anaesthetic techniques were used; thiopentone-enflurane-nitrous oxide and thiopentone-fentanyl-nitrous oxide. The patients were comparable in age, weight, length of anaesthesia and time for recovery to street fitness after anaesthesia. The amount of thiopentone administered to the thiopentone-fentanyl-nitrous oxide groups was significantly greater than that administered to the thiopentone-enflurane-nitrous oxide groups. It is concluded that premedication with meperidine and atropine did not significantly prolong recovery to street fitness after out-patient surgery. Fear of prolonged recovery should not affect the decision to use premedication.

Adult↗
PubMed1981

Evidence that rnmB is the operator of the Escherichia coli recA gene.

rnmB281 leads to high constitutive levels of recA protein such that no increase after UV-inducing treatment occurs. The mutation maps in or near the portion of recA corresponding to the NH2-terminal end of the protein. Examination of the recA proteins from rnmB+ recA-/rnmB281 recA+ heterozygotes suggests that both rnmB alleles are cis-acting and codominant. This is the behavior expected from alleles of a regulatory gene such as an operator or promoter of recA. The possibility that rnmB mutations occur in the promoter of recA. The possibility that rnmB mutations occur in the promoter of recA, though not ruled out, seems unlikely based on the structure of the regulatory region of recA. This suggests that rnmB mutations are operator constitutive mutations of the recA gene and should be called recAo mutations. The UV-irradiation responses of recAo+ and recAo281 strains, both recA+, are compared and inferences are drawn about the roles of large amounts of recA protein in producing the responses.

Bacterial Proteins↗
PubMed1981

Genetic analysis of the RecE pathway of genetic recombination in Escherichia coli K-12.

The RecE pathway of genetic recombination in Escherichia coli K-12 was defined to be the pathway that is utilized in deoxyribonucleic acid exonuclease V (ExoV)-defective cells which express constitutively recE+, the structural gene for deoxyribonucleic acid exonuclease VIII. Dependence on ExoVIII was shown by the occurrence in a recB21 sbcA23 strain of recombination deficiency mutations in recE, the structural gene for ExoVIII. Point mutations in recE were found as well as deletion mutations in which the entire Rac prophage, carrying recE, was lost. In addition, strain construction and mutagenesis revealed the dependence of the RecE pathway on recA+ and on recF+. Dependence on a fourth gene was shown by a mutation (rec-77) which does not map near the other genes. The problem of distinguishing the RecE pathway from that previously called RecF is discussed.

Escherichia coli↗
PubMed1981

Overproduction of the Escherichia coli recA protein without stimulation of its proteolytic activity.

Plasmid pBEU14, which carries the Escherichia coli recA+ gene and which can be amplified by manipulation of growth temperature, was constructed. When pBEU14 deoxyribonucleic acid was amplified, a high rate of synthesis and accumulation of recA protein resulted. Amplification of the recA gene and protein did not cause induction of prophage lambda, indicating that the proteolytic activity of the recA protein was not stimulated.

Bacterial Proteins↗