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Human gene mapping using the polymerase chain reaction / Malika Nebbali 

Public ISBD Titre : Human gene mapping using the polymerase chain reaction Type de document : texte imprimé Auteurs : Malika Nebbali, Auteur ; Stuart Brown, Directeur de thèse Editeur : Nottingham : University of Nottingham Année de publication : 1992 Importance : 153 f. Présentation : ill. Format : 30 cm. Note générale : Thèse de Doctorat : Génie Chimique : Angleterre, University of Nottingham : 1992 Bibliogr. f. 154 - 167 . Annexe [3] f Langues : Anglais (eng) Mots-clés : Gene  mapping Hybridisation  techniques DNA  polymorphism Polymerase  chain  reaction  methods Chemical  carcinogenesis Chromatographic  techniques Immunological  methods Index. décimale : D001492 Résumé : There has been strong association between cigarette smoking and the incidence of lung cancer. Cigarette smoke is a rich source of PAH and it has been reported that high AHH activities in individuals makes them at high risk to lung cancer if they smoke cigarettes. In previous work a gene necessary for AHH induction was found to segregate with chromosome 2 markers; MDH-1, IDH-1 and ACP in human x mouse hybrid cells and was mapped to 2q31 --> 2pter. However a hybrid subclone containing a small deletion 2p25 --> 2pter which did not express MDH-1, ACP-1 or AHH was isolated. This could mean that the genes for these enzymes were actually missing from the chromosome and thus are located in the small deleted region. It is also possible that they are present elsewhere on the chromosome byt in an inactive form. Whichever explanation was true mapping of the three enzymes was of interest and MDH-1 was chosen as the enzyme to be mapped. Mapping of MDH-1 would strongly suggest that the genes for the other two enzymes were located in the same region. Since no data was available concerning the human MDH-1 gene, a strategy was designed in which MDH-1 was to be purified to homogeneity from red blood cells and the purified protein was to used to raise antibodies in rabbits. The aim then was to isolate the MDH-1 gene by immunological screening of a human λgt11 cDNA expression library. The gene would be subsquently used as a probe in in-situ hybridization techniques. MDH-1 was purified from human red blood cells using polyethylene glycol fractionation. The activity precipitated between 8 and 16%. Solubilised protein was separated by DEAE-cellulose and agarose affinity chromatography. Homogeneous enzyme was prepared by removing the contaminating proteins by FPLC using a Superose 12 gel filtration column. A 5500 fold purification was achieved. The purified preparation, had a molecular weight of approximatly 76000 and 38000 after sodium dodecyl sulphate gel electrophoresis. Thus, the enzyme is composed of two subunits of equal molecular weight. Antibodies were raised by injecting a rabbit with the agarose affinity fraction, but failed to detect the gene of interest after immunological screening of a human placental cDNA expression library. A second strategy was designed which involved sequencing peptides generated from the pure protein. The data obtained was used to sythesize oligonucleotides which served as primers for the amplification of specific regions of DNA from the MDH-1 gene via the polymerase chain reaction (PCR). Four peptides were produced by cyanogen bromide cleavage, purified using HPLC and sesuenced. It was found that the enzyme is highly homologous to the mouse enzyme as only one major difference was apparent at residue 288 where leucine was replaced by phenyalanine in humans. Three oligonucleotides called, 21, 31 and 30 mers respectively were synthesized P₄ and P₅ were used in PCRs for the amplification of specific regions of the MDH-1 gene which corresponded to 872 bp and 708 bp in length for the mouse and human genomic DNA respectively. The 708 bp product was purified from the agarose gel and was subjected to direct DNA sequencing. Comparison of the data obtained with the known mouse MDH-1 gene sequence strongly suggested the 708 bp band being amplified was human MDH-1. The fact that the amplification of human and mouse genomic DNA yielded products of different size had a major impact on future work. It was realised that it would not be necessary to carry out in-situ hybridisation to dorectly test for the presence or absence of the MDH-1 gene in hybrid cells. PCR alone would be able to give the desired results in a fraction of the time. Human gene mapping using the polymerase chain reaction [texte imprimé] / Malika Nebbali, Auteur ; Stuart Brown, Directeur de thèse . - Nottingham : University of Nottingham, 1992 . - 153 f. : ill. ; 30 cm. Thèse de Doctorat : Génie Chimique : Angleterre, University of Nottingham : 1992 Bibliogr. f. 154 - 167 . Annexe [3] f Langues : Anglais (eng) Mots-clés : Gene  mapping Hybridisation  techniques DNA  polymorphism Polymerase  chain  reaction  methods Chemical  carcinogenesis Chromatographic  techniques Immunological  methods Index. décimale : D001492 Résumé : There has been strong association between cigarette smoking and the incidence of lung cancer. Cigarette smoke is a rich source of PAH and it has been reported that high AHH activities in individuals makes them at high risk to lung cancer if they smoke cigarettes. In previous work a gene necessary for AHH induction was found to segregate with chromosome 2 markers; MDH-1, IDH-1 and ACP in human x mouse hybrid cells and was mapped to 2q31 --> 2pter. However a hybrid subclone containing a small deletion 2p25 --> 2pter which did not express MDH-1, ACP-1 or AHH was isolated. This could mean that the genes for these enzymes were actually missing from the chromosome and thus are located in the small deleted region. It is also possible that they are present elsewhere on the chromosome byt in an inactive form. Whichever explanation was true mapping of the three enzymes was of interest and MDH-1 was chosen as the enzyme to be mapped. Mapping of MDH-1 would strongly suggest that the genes for the other two enzymes were located in the same region. Since no data was available concerning the human MDH-1 gene, a strategy was designed in which MDH-1 was to be purified to homogeneity from red blood cells and the purified protein was to used to raise antibodies in rabbits. The aim then was to isolate the MDH-1 gene by immunological screening of a human λgt11 cDNA expression library. The gene would be subsquently used as a probe in in-situ hybridization techniques. MDH-1 was purified from human red blood cells using polyethylene glycol fractionation. The activity precipitated between 8 and 16%. Solubilised protein was separated by DEAE-cellulose and agarose affinity chromatography. Homogeneous enzyme was prepared by removing the contaminating proteins by FPLC using a Superose 12 gel filtration column. A 5500 fold purification was achieved. The purified preparation, had a molecular weight of approximatly 76000 and 38000 after sodium dodecyl sulphate gel electrophoresis. Thus, the enzyme is composed of two subunits of equal molecular weight. Antibodies were raised by injecting a rabbit with the agarose affinity fraction, but failed to detect the gene of interest after immunological screening of a human placental cDNA expression library. A second strategy was designed which involved sequencing peptides generated from the pure protein. The data obtained was used to sythesize oligonucleotides which served as primers for the amplification of specific regions of DNA from the MDH-1 gene via the polymerase chain reaction (PCR). Four peptides were produced by cyanogen bromide cleavage, purified using HPLC and sesuenced. It was found that the enzyme is highly homologous to the mouse enzyme as only one major difference was apparent at residue 288 where leucine was replaced by phenyalanine in humans. Three oligonucleotides called, 21, 31 and 30 mers respectively were synthesized P₄ and P₅ were used in PCRs for the amplification of specific regions of the MDH-1 gene which corresponded to 872 bp and 708 bp in length for the mouse and human genomic DNA respectively. The 708 bp product was purified from the agarose gel and was subjected to direct DNA sequencing. Comparison of the data obtained with the known mouse MDH-1 gene sequence strongly suggested the 708 bp band being amplified was human MDH-1. The fact that the amplification of human and mouse genomic DNA yielded products of different size had a major impact on future work. It was realised that it would not be necessary to carry out in-situ hybridisation to dorectly test for the presence or absence of the MDH-1 gene in hybrid cells. PCR alone would be able to give the desired results in a fraction of the time.

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Documents numériques

NEBBALI.Malika.pdf URL

Optical local area network feturing a variable output power transmitter / A. Boudghene Stambouli 

Public ISBD Titre : Optical local area network feturing a variable output power transmitter Type de document : texte imprimé Auteurs : A. Boudghene Stambouli, Auteur ; A. J. Lowery, Directeur de thèse Editeur : Nottingham : University of Nottingham Année de publication : 1985 Importance : 79 f. Présentation : ill. Format : 27 cm. Note générale : Mémoire de Magister : Électronique : Nottingham, University of Nottingham : 1985 Annexe [12] f. Bibliogr. f. 84 Langues : Anglais (eng) Mots-clés : Computer  --  networks  ;  Copper  --  conductors  ;  Optical  --  fibre  --  network  ;  Ethernet Index. décimale : M007185 Résumé : Local computer networks which communicate over copper conductors have been developed both to promote resource sharing and provide increased performance. in this report the use of fibre optics in such networks is considered, and a status report on the optical fibre network experiment which operates at 1Mbit/s and 10Mbit/s is given. The system performance of the Optical Ethernet experiment, which uses an LED with power output control possibility, multimode step index fibre, PIN photodiode, and incorporating a collision-detection-device is described. Circuitry developed for each sub-system is presented with its characteristic curves and output waveforms for a specific input signal. The performance of the receiver used is examined, the noise present in the unit, the minimum power required are measured as well as the value of its sensitivity for a SNR of 21.6dBm which yields a BER of 10⁻⁹ . Finally suggested circuit for measurement of the BER is presented as an expansion possibility for this project. Optical local area network feturing a variable output power transmitter [texte imprimé] / A. Boudghene Stambouli, Auteur ; A. J. Lowery, Directeur de thèse . - Nottingham : University of Nottingham, 1985 . - 79 f. : ill. ; 27 cm. Mémoire de Magister : Électronique : Nottingham, University of Nottingham : 1985 Annexe [12] f. Bibliogr. f. 84 Langues : Anglais (eng) Mots-clés : Computer  --  networks  ;  Copper  --  conductors  ;  Optical  --  fibre  --  network  ;  Ethernet Index. décimale : M007185 Résumé : Local computer networks which communicate over copper conductors have been developed both to promote resource sharing and provide increased performance. in this report the use of fibre optics in such networks is considered, and a status report on the optical fibre network experiment which operates at 1Mbit/s and 10Mbit/s is given. The system performance of the Optical Ethernet experiment, which uses an LED with power output control possibility, multimode step index fibre, PIN photodiode, and incorporating a collision-detection-device is described. Circuitry developed for each sub-system is presented with its characteristic curves and output waveforms for a specific input signal. The performance of the receiver used is examined, the noise present in the unit, the minimum power required are measured as well as the value of its sensitivity for a SNR of 21.6dBm which yields a BER of 10⁻⁹ . Finally suggested circuit for measurement of the BER is presented as an expansion possibility for this project.

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M007185	M007185	Papier	Bibliothèque centrale	Mémoire de Magister	Disponible

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BOUDGHENE-STAMBOULI.A.pdf URL

Microbial responses to surfactants / Leila Laouar 

Public ISBD Titre : Microbial responses to surfactants Type de document : texte imprimé Auteurs : Leila Laouar, Auteur ; K. C. Lowe, Directeur de thèse ; B. J. Mulligan, Directeur de thèse Editeur : Nottingham : University of Nottingham Année de publication : 1993 Importance : 186 f. Présentation : ill. Format : 30 cm. Note générale : Thèse de Doctorat : Génie Chimique : Angleterre, University of Nottingham : 1993 Annexe f. 187 - 226 . Bibliogr. f. 227 - 248 Langues : Anglais (eng) Mots-clés : Pluronic  F-68 Microbial  cells Saccharomyces  cerevisiae Triton  X-100 Surfactants Alcohol  dehydrogenase Index. décimale : D000893 Résumé : The effects of commercial grade pluronic F-68 or its purified fraction or Triton X-100 on growth of microbial cells in culture have been studied. Growth of Saccharomyces cerevisiae, as determined by viable cell counts, was unaffected by culture with 0.1 - 10.0% (W/V) of commercial grade Pluronic F-68 or its purified fraction. However, cultures treated with 10.0% (W/V) commercial grade or purified Pluronic F-68 showed significabtly reduced absorbance up to the onset of the stationary phase of growth, as compared to controls. The responses to Pluronic F-68 varied according to concentration, age and purity of surfactant. Additionally, the storage temperature influenced the properties of Pluronic F-68. Growth in the presence of up to 10.0% (W/V) commercial or purified Pluronic F-68 had no effect on cell-associated protein concentration. However, a slight, but reproducible, increase in protein concentration was observed during the exponential/stationary phase in cultures supplemented with 0.1 - 1.0% Pluronic F-68. In addition, whole cell polypeptide profiles were unaffected by the presence of pluronic F-68. Triton X-100 altered growth kinetics, as reflected by an extended stationary phase and a slowing of the senescence phase. Additionally, in cultures supplemented with Triton, cell-associated protein concentration was proportional to cellular growth. Culture in the presence of Triton X-100 did not alter the polypeptide profiles characteristic of untreated cells at different phases of growth. The application of surfactants for cell permeabilization was also studied using both Pluronic F-68 and Triton X-100, together with Pluronic F-38 and pluronic L-35. Permeabilization was assayed by direct measurement of alcohol dehydrogenase (ADH) activity in whole cells. Mean ADH activity following incubation of S. cerevisiae with 0.2% (W/V or V/V) of either commercial or purified Pluronic F-68, Pluronic F-38, Pluronic L-35 or Triton X-100 for 30 min at 20°C was significantly greater than when cells were exposed to an identical concentration of the previously studied permeabilizing agent, cetyltrimethylammonium bromide (CTAB). Microbial responses to surfactants [texte imprimé] / Leila Laouar, Auteur ; K. C. Lowe, Directeur de thèse ; B. J. Mulligan, Directeur de thèse . - Nottingham : University of Nottingham, 1993 . - 186 f. : ill. ; 30 cm. Thèse de Doctorat : Génie Chimique : Angleterre, University of Nottingham : 1993 Annexe f. 187 - 226 . Bibliogr. f. 227 - 248 Langues : Anglais (eng) Mots-clés : Pluronic  F-68 Microbial  cells Saccharomyces  cerevisiae Triton  X-100 Surfactants Alcohol  dehydrogenase Index. décimale : D000893 Résumé : The effects of commercial grade pluronic F-68 or its purified fraction or Triton X-100 on growth of microbial cells in culture have been studied. Growth of Saccharomyces cerevisiae, as determined by viable cell counts, was unaffected by culture with 0.1 - 10.0% (W/V) of commercial grade Pluronic F-68 or its purified fraction. However, cultures treated with 10.0% (W/V) commercial grade or purified Pluronic F-68 showed significabtly reduced absorbance up to the onset of the stationary phase of growth, as compared to controls. The responses to Pluronic F-68 varied according to concentration, age and purity of surfactant. Additionally, the storage temperature influenced the properties of Pluronic F-68. Growth in the presence of up to 10.0% (W/V) commercial or purified Pluronic F-68 had no effect on cell-associated protein concentration. However, a slight, but reproducible, increase in protein concentration was observed during the exponential/stationary phase in cultures supplemented with 0.1 - 1.0% Pluronic F-68. In addition, whole cell polypeptide profiles were unaffected by the presence of pluronic F-68. Triton X-100 altered growth kinetics, as reflected by an extended stationary phase and a slowing of the senescence phase. Additionally, in cultures supplemented with Triton, cell-associated protein concentration was proportional to cellular growth. Culture in the presence of Triton X-100 did not alter the polypeptide profiles characteristic of untreated cells at different phases of growth. The application of surfactants for cell permeabilization was also studied using both Pluronic F-68 and Triton X-100, together with Pluronic F-38 and pluronic L-35. Permeabilization was assayed by direct measurement of alcohol dehydrogenase (ADH) activity in whole cells. Mean ADH activity following incubation of S. cerevisiae with 0.2% (W/V or V/V) of either commercial or purified Pluronic F-68, Pluronic F-38, Pluronic L-35 or Triton X-100 for 30 min at 20°C was significantly greater than when cells were exposed to an identical concentration of the previously studied permeabilizing agent, cetyltrimethylammonium bromide (CTAB).

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Documents numériques

LAOUAR.Leila.pdf URL

Experimental data plotter / Boubakeur Benzeltout 

Public ISBD Titre : Experimental data plotter Type de document : texte imprimé Auteurs : Boubakeur Benzeltout, Auteur ; B. Wilson, Directeur de thèse Editeur : Nottingham : University of Nottingham Année de publication : 1984 Importance : 80 f. Présentation : ill. Format : 27 cm. Note générale : Mémoire de Master : Electronique : Nottingham, University of Nottingham : 1984 Annexe f. 81 - 93 . Bibliogr. f. 94 Langues : Anglais (eng) Mots-clés : Microcomputer Analogue  X-Y  recorder Cubic  splines Polynominal  interpolation Index. décimale : Ms00284 Résumé : The design of an interface between a microcomputer and an analogue X-Y recorder is described in detail. Given a set of experimental data points the line of best fit is generated by a Fortran routine using cubic splines to calculate the intermediate values. An assembly language routine is used to draw the axes, scale them, and then to draw the curve using the previously calculated intermediate values. Comments on both routines and their use are included. Experimental data plotter [texte imprimé] / Boubakeur Benzeltout, Auteur ; B. Wilson, Directeur de thèse . - Nottingham : University of Nottingham, 1984 . - 80 f. : ill. ; 27 cm. Mémoire de Master : Electronique : Nottingham, University of Nottingham : 1984 Annexe f. 81 - 93 . Bibliogr. f. 94 Langues : Anglais (eng) Mots-clés : Microcomputer Analogue  X-Y  recorder Cubic  splines Polynominal  interpolation Index. décimale : Ms00284 Résumé : The design of an interface between a microcomputer and an analogue X-Y recorder is described in detail. Given a set of experimental data points the line of best fit is generated by a Fortran routine using cubic splines to calculate the intermediate values. An assembly language routine is used to draw the axes, scale them, and then to draw the curve using the previously calculated intermediate values. Comments on both routines and their use are included.

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Documents numériques

BENZELTOUT.Boubakeur.pdf URL

Analysis of the tapered waveguide / A. Belghoraf 

Public ISBD Titre : Analysis of the tapered waveguide Type de document : texte imprimé Auteurs : A. Belghoraf, Auteur ; R. L. Beurle, Directeur de thèse Editeur : Nottingham : University of Nottingham Année de publication : 1984 Importance : 110 f. Présentation : ill. Format : 27 cm. Note générale : Thèse de Doctorat : Électronique : Angleterre, University of Nottingham : 1984 Bibliogr. f. 130 - 132. Annexe f. 111 - 129 Langues : Anglais (eng) Mots-clés : Tapered  waveguide  problems Plane  wave  spectral  analysis Intrinsic  mode  theory Parabolic  equation  method Index. décimale : D003484 Résumé : This thesis describes analytical and numerical investigations of tapered waveguide problems, for intergrated optics applications. A plane wave spectral analysis, models the propagation process of the tapered waveguide and introduces the concept of an Intrinsic spectral Integral, which turns out to be in good agreement with calculation in terms of Adiabatic modes. This allows us to extend the Intrinsic mode concept beyond the singularity where the Adiabatic mode concept breaks down. In this sense, the implementation of the resulting spectral formulation, for the case of homogeneous media, contains all information pertinent to the modal propagation mechanism, inside and outside the tapered waveguide; before and after the singularity caused by cut off of the Adiabatic mode. The thesis is mainly concerned with implementing the Intrinsic mode theory as a numerical computational tool. In this respect, very good agreement is demonstrated between this model and calculations performed numerically using the parabolic equation method. On the other hand, the new model contains far greater physical and analytical possibilities than previous methods. Analysis of the tapered waveguide [texte imprimé] / A. Belghoraf, Auteur ; R. L. Beurle, Directeur de thèse . - Nottingham : University of Nottingham, 1984 . - 110 f. : ill. ; 27 cm. Thèse de Doctorat : Électronique : Angleterre, University of Nottingham : 1984 Bibliogr. f. 130 - 132. Annexe f. 111 - 129 Langues : Anglais (eng) Mots-clés : Tapered  waveguide  problems Plane  wave  spectral  analysis Intrinsic  mode  theory Parabolic  equation  method Index. décimale : D003484 Résumé : This thesis describes analytical and numerical investigations of tapered waveguide problems, for intergrated optics applications. A plane wave spectral analysis, models the propagation process of the tapered waveguide and introduces the concept of an Intrinsic spectral Integral, which turns out to be in good agreement with calculation in terms of Adiabatic modes. This allows us to extend the Intrinsic mode concept beyond the singularity where the Adiabatic mode concept breaks down. In this sense, the implementation of the resulting spectral formulation, for the case of homogeneous media, contains all information pertinent to the modal propagation mechanism, inside and outside the tapered waveguide; before and after the singularity caused by cut off of the Adiabatic mode. The thesis is mainly concerned with implementing the Intrinsic mode theory as a numerical computational tool. In this respect, very good agreement is demonstrated between this model and calculations performed numerically using the parabolic equation method. On the other hand, the new model contains far greater physical and analytical possibilities than previous methods.

Exemplaires

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D003484	D003484	Papier + ressource électronique	Bibliothèque centrale	Thèse de Doctorat	Disponible	Electronique	Consultation sur place/Téléchargeable

Documents numériques

BELGHORAF.A.pdf URL

Discharge coefficient of film cooling holes / Saad Benmansour 

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Portable equipment for monitoring and recording human postures / B. Mehenni 

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A speech message store and readout system using a delta modulation technique / Meliani, Hamza 

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An expert system approach to robot RIG controller design / Boubakeur Benzeltout

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