DOCSLIB.ORG

Regulation of Acetyl Phosphate-Dependent Acetylation and Identification of Novel Lysine Acetyltransferases in Escherichia Coli

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Regulation of Acetyl Phosphate-Dependent Acetylation and Identification of Novel Lysine Acetyltransferases in Escherichia Coli Loyola University Chicago Loyola eCommons Dissertations Theses and Dissertations 2019 Regulation of Acetyl Phosphate-Dependent Acetylation and Identification of Novel Lysine Acetyltransferases in Escherichia Coli David George Christensen Follow this and additional works at: https://ecommons.luc.edu/luc_diss Part of the Microbiology Commons Recommended Citation Christensen, David George, "Regulation of Acetyl Phosphate-Dependent Acetylation and Identification of Novel Lysine Acetyltransferases in Escherichia Coli" (2019). Dissertations. 3328. https://ecommons.luc.edu/luc_diss/3328 This Dissertation is brought to you for free and open access by the Theses and Dissertations at Loyola eCommons. It has been accepted for inclusion in Dissertations by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License. Copyright © 2019 David George Christensen LOYOLA UNIVERSITY CHICAGO REGULATION OF ACETYL PHOSPHATE-DEPENDENT ACETYLATION AND IDENTIFICATION OF NOVEL LYSINE ACETYLTRANSFERASES IN ESCHERICHIA COLI A DISSERTATION SUBMITTED TO THE FACULTY OF THE GRADUATE SCHOOL IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY PROGRAM IN MICROBIOLOGY AND IMMUNOLOGY BY DAVID GEORGE CHRISTENSEN CHICAGO, ILLINOIS MARCH 2019 Copyright by David G. Christensen, 2019 All rights reserved. ACKNOWLEDGMENTS I must first thank my parents for their unending love and encouragement throughout my education that has brought me to this point. I would also like to express my thanks to my Aunt Heidi who has greatly supported my education and ensured that I had many opportunities. I would also like to thank my future spouse, Carolyn, for constant love, support, and understanding. I would like to thank my mentor, Dr. Alan Wolfe, who has given me guidance and support that has greatly fostered my growth as a scientist. I would like to thank the Wolfe lab members past and present who have shaped a wonderful community for scientific and non- scientific discussions, especially Drs. Robert Davis, Linda Hu, and Bruno Lima upon whose work my studies stand. I am thankful for each of the students that I mentored and who taught me as much as I taught them. I would like to thank the Microbiology and Immunology Department for being a brilliant group who have provided diverse perspectives on my work. I would also like to thank my committee for giving me constructive feedback on my work at the bench, in my writing, and in my thinking. I would like to thank my collaborators who have provided me with valuable expertise and ideas that were essential to drive my studies. I’d like to especially thank Dr. Christopher Rao and James Orr for their assistance in measuring metabolites, Dr. Birgit Schilling for her assistance in performing mass spectrometry, and Dr. Misty Kuhn for her assistance with enzymology. iii For my parents who have provided me with every opportunity and Carolyn who provides me with love, encouragement, and joy. You are an aperture through which the universe is looking at and exploring itself. Alan W. Watts . TABLE OF CONTENTS ACKNOWLEDGMENTS ............................................................................................................. iii LIST OF TABLES .......................................................................................................................... x LIST OF FIGURES ...................................................................................................................... xi LIST OF ABBREVIATIONS ..................................................................................................... xiv ABSTRACT ...................................................................................................................................xv CHAPTER ONE: LITERATURE REVIEW ...................................................................................1 Introduction ...................................................................................................................................1 Acetylation ....................................................................................................................................3 Acetylation – From Small Molecules to Proteins ......................................................................3 Nε-Lysine Acetylation ...............................................................................................................5 Acetyltransferases in Bacteria ..................................................................................................12 YfiQ, the Only Known Acetyltransferase in E. coli and S. enterica .......................................16 Deacetylases in Bacteria ..........................................................................................................19 CobB, the Only Known Deacetylase in E. coli ........................................................................20 Effects of Enzymatic Acetylation and Deacetylation on Physiology and Pathogenesis .........29 Glucose-Induced Acetylation in E. coli Is Acetyl Phosphate-Dependent ...............................33 Non-Enzymatic Acetylation via AcP and AcCoA ...................................................................34 A Simplified Overview of Central Metabolism and Overflow Metabolism...............................38 Stationary Phase Cultures of E. coli – Physiological Shifts .......................................................43 Summary .....................................................................................................................................46 CHAPTER TWO: MATERIALS AND METHODS ....................................................................48 Bacterial Strains, Plasmids, and Primers ....................................................................................48 Culture Conditions ......................................................................................................................48 Generalized P1 Transduction ......................................................................................................57 Elimination of Kanamycin Cassettes Flanked by FRT Sites ......................................................58 Site-Directed Mutagenesis ..........................................................................................................58 FRUIT: Flexible Recombineering Using Integration of thyA.....................................................58 Transformation ............................................................................................................................59 TBF ..........................................................................................................................................59 Electroporation .........................................................................................................................60 Measurement of Glucose and Acetate in Media .........................................................................60 Western Immunoblot Analysis ...................................................................................................61 Isolation and Qualitative Assessment of Protein Aggregates .....................................................62 Carbonylation Analysis ...............................................................................................................63 Purification of His-Tagged Proteins ...........................................................................................64 In Vitro Acetylation Assays ........................................................................................................64 Enzymatic ................................................................................................................................64 vi Non-Enzymatic ........................................................................................................................65 Starvation Survival Assay ...........................................................................................................65 DNA Alkylation Survival Assay ................................................................................................65 Mucoidy Assay ...........................................................................................................................66 Motility Assay .............................................................................................................................66 Mass Spectrometry......................................................................................................................66 Preparing Samples ...................................................................................................................66 Proteolytic Digestion of Protein Lysates and Enrichment of Acetylated Peptides ..................67 Mass Spectrometric Analysis ...................................................................................................68 CHAPTER THREE: EXPERIMENTAL RESULTS ....................................................................70 Characterizing the Timing and Dynamics of Glucose-Dependent Acetylation..........................70 Introduction ..............................................................................................................................70 Carbon-Induced, AcP-Dependent Acetylation Accumulates during Stationary Phase ...........71 Glucose Is Not Consumed until Stationary Phase in TB7 .......................................................74
Load more

Recommended publications
  • Is There Any Role for Camp–CRP in Carbon Catabolite Repression of The
    LINK TO ORIGINAL ARTICLE CORRESPONDENCE LINK TO Author’S REPLY glucose. It has been established that the level of cAMP in lactose-grown cells is low Is there any role for cAMP–CRP compared with other less-preferred carbon sources, but nevertheless is slightly higher in carbon catabolite repression of than the level of cAMP in glucose-grown cells9. Indeed, phosphorylated enzyme the Escherichia coli lac operon? IIAglc (which activates adenylate cyclase) is detected in lactose-grown cells but not Martine Crasnier-Mednansky in glucose-grown cells, as supported by the phosphorylation state of enzyme IIAglc In their review on carbon catabolite repres- case, then adding cAMP should have only detected by Western blotting10. sion (CCR) in bacteria, Boris Görke and a limited effect on diauxie or at least should Finally, other data suggest a role for Jörg Stülke (Carbon catabolite repression stimulate inducer exclusion by enhancing cAMP–CRP in diauxie. For example, a in bacteria: many ways to make the most glucose transport. Consequently, a further PTS sugar-like mannitol can substitute for out of nutrients. Nature Rev. Microbiol. 6, decrease in β-galactosidase expression in the glucose in diauxie production in E. coli 613–624 (2008))1 specifically analysed CCR presence of cAMP would result in enhance- whereas others, such as fructose, do not2. in Escherichia coli. The authors reported their ment of the diauxic lag. In any case, adding This correlates with the observation that view that inducer exclusion, by blocking cAMP would eliminate the diauxic lag. the level of cAMP in mannitol-grown cells the entry of lactose, is the main contributor The next contentious issue arises from the is in the same range as in glucose-grown to CCR when wild-type E.
    [Show full text]
  • VU Research Portal
    VU Research Portal Dynamic regulation of yeast glycolysis through trehalose cycling van Heerden, J.H. 2016 document version Publisher's PDF, also known as Version of record Link to publication in VU Research Portal citation for published version (APA) van Heerden, J. H. (2016). Dynamic regulation of yeast glycolysis through trehalose cycling: a probabilistic view of metabolic transitions. General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. E-mail address: [email protected] Download date: 09. Oct. 2021 Dynamic regulation of yeast glycolysis through trehalose cycling a probabilistic view of metabolic transitions Johan H. van Heerden Members of the Doctoral Examination Committee: prof. dr. Hans V. Westerhoff VU University, Amsterdam prof. dr. Stefan Hohmann University of Gothenburg prof. dr. Barbara
    [Show full text]
  • Camp Receptor Protein–Camp Plays a Crucial Role in Glucose–Lactose Diauxie by Activating the Major Glucose Transporter Gene in Escherichia Coli
    Proc. Natl. Acad. Sci. USA Vol. 94, pp. 12914–12919, November 1997 Biochemistry cAMP receptor protein–cAMP plays a crucial role in glucose–lactose diauxie by activating the major glucose transporter gene in Escherichia coli KEIKO KIMATA*, HIDEYUKI TAKAHASHI*, TOSHIFUMI INADA*, PIETER POSTMA†, AND HIROJI AIBA*‡ *Department of Molecular Biology, School of Science, Nagoya University, Chikusa, Nagoya 464-01, Japan; and †E. C. Slater Institute, BioCentrum, University of Amsterdam, Plantage Muidergracht 12, 1018 TV Amsterdam, The Netherlands Communicated by Sydney Kustu, University of California, Berkeley, CA, September 17, 1997 (received for review May 2, 1997) ABSTRACT The inhibition of b-galactosidase expression certain conditions, for example, when added to cells growing in a medium containing both glucose and lactose is a typical on a poor carbon source such as glycerol or succinate (6, 7). example of the glucose effect in Escherichia coli. We studied the Glucose is thought to reduce cAMP level by decreasing the glucose effect in the lacL8UV5 promoter mutant, which is phosphorylated form of enzyme IIAGlc, which is proposed to independent of cAMP and cAMP receptor protein (CRP). A be involved in the activation of adenylate cyclase (3–5). strong inhibition of b-galactosidase expression by glucose and Glucose also is known to reduce the CRP level through the a diauxic growth were observed when the lacL8UV5 cells were autoregulation of the crp gene (7–10). grown on a glucose–lactose medium. The addition of isopropyl When E. coli finds both glucose and lactose in the medium, b-D-thiogalactoside to the culture medium eliminated the it preferentially uses the glucose, and the use of lactose is glucose effect.
    [Show full text]
  • Coupling Gene Regulatory Patterns to Bioprocess Conditions to Optimize
    Peng et al. Biotechnol Biofuels (2017) 10:43 DOI 10.1186/s13068-017-0728-x Biotechnology for Biofuels RESEARCH Open Access Coupling gene regulatory patterns to bioprocess conditions to optimize synthetic metabolic modules for improved sesquiterpene production in yeast Bingyin Peng1, Manuel R. Plan1,2, Alexander Carpenter1, Lars K. Nielsen1 and Claudia E. Vickers1* Abstract Background: Assembly of heterologous metabolic pathways is commonly required to generate microbial cell facto- ries for industrial production of both commodity chemicals (including biofuels) and high-value chemicals. Promoter- mediated transcriptional regulation coordinates the expression of the individual components of these heterologous pathways. Expression patterns vary during culture as conditions change, and this can influence yeast physiology and productivity in both positive and negative ways. Well-characterized strategies are required for matching transcrip- tional regulation with desired output across changing culture conditions. Results: Here, constitutive and inducible regulatory mechanisms were examined to optimize synthetic isoprenoid metabolic pathway modules for production of trans-nerolidol, an acyclic sesquiterpene alcohol, in yeast. The choice of regulatory system significantly affected physiological features (growth and productivity) over batch cultivation. Use of constitutive promoters resulted in poor growth during the exponential phase. Delaying expression of the assembled metabolic modules using the copper-inducible CUP1 promoter resulted in a 1.6-fold increase in the exponential- phase growth rate and a twofold increase in productivity in the post-exponential phase. However, repeated use of the CUP1 promoter in multiple expression cassettes resulted in genetic instability. A diauxie-inducible expression system, based on an engineered GAL regulatory circuit and a set of four different GAL promoters, was characterized and employed to assemble nerolidol synthetic metabolic modules.
    [Show full text]
  • Bacterial Gene Regulation in Diauxic and Non-Diauxic Growth
    ARTICLE IN PRESS Journal of Theoretical Biology 244 (2007) 326–348 www.elsevier.com/locate/yjtbi Bacterial gene regulation in diauxic and non-diauxic growth Atul Naranga,Ã, Sergei S. Pilyuginb aDepartment of Chemical Engineering, University of Florida, Gainesville, FL 32611-6005, USA bDepartment of Mathematics, University of Florida, Gainesville, FL 32611-8105, USA Received 18 May 2006; received in revised form 24 July 2006; accepted 9 August 2006 Available online 12 August 2006 Abstract When bacteria are grown in a batch culture containing a mixture of two growth-limiting substrates, they exhibit a rich spectrum of substrate consumption patterns including diauxic growth, simultaneous consumption, and bistable growth. In previous work, we showed that a minimal model accounting only for enzyme induction and dilution captures all the substrate consumption patterns [Narang, A., 1998a. The dynamical analogy between microbial growth on mixtures of substrates and population growth of competing species. Biotechnol. Bioeng. 59, 116–121, Narang, A., 2006. Comparitive analysis of some models of gene regulation in mixed-substrate microbial growth, J. Theor. Biol. 242, 489–501]. In this work, we construct the bifurcation diagram of the minimal model, which shows the substrate consumption pattern at any given set of parameter values. The bifurcation diagram explains several general properties of mixed-substrate growth. (1) In almost all the cases of diauxic growth, the ‘‘preferred’’ substrate is the one that, by itself, supports a higher specific growth rate. In the literature, this property is often attributed to the optimality of regulatory mechanisms. Here, we show that the minimal model, which accounts for induction and growth only, displays the property under fairly general conditions.
    [Show full text]
  • Growth Strategy of Microbes on Mixed Carbon Sources
    Growth strategy of microbes on mixed carbon sources Xin Wang1,2,†, Kang Xia1,3,†, Xiaojing Yang1, Chao Tang1,* 1Center for Quantitative Biology, School of Physics and Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China. 2Channing Division of Network Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA. 3College of Life Sciences, Wuhan University, Wuhan 430072, China. †These authors contributed equally to this work. *Correspondence and requests for materials should be addressed to C.T. (email: [email protected]). Keywords: Cell growth, diauxie, co-utilization of carbon sources, optimality, metabolic network, catabolite repression, amino acids synthesis 1 Abstract A classic problem in microbiology is that bacteria display two types of growth behavior when cultured on a mixture of two carbon sources: the two sources are sequentially consumed one after another (diauxie) or they are simultaneously consumed (co-utilization). The search for the molecular mechanism of diauxie led to the discovery of the lac operon. However, questions remain as why microbes would bother to have different strategies of taking up nutrients. Here we show that diauxie versus co-utilization can be understood from the topological features of the metabolic network. A model of optimal allocation of protein resources quantitatively explains why and how the cell makes the choice. In case of co-utilization, the model predicts the percentage of each carbon source in supplying the amino acid pools, which is quantitatively verified by experiments. Our work solves a long-standing puzzle and provides a quantitative framework for the carbon source utilization of microbes.
    [Show full text]
  • Emergence of Diauxie As an Optimal Growth Strategy Under Resource Allocation Constraints in Cellular Metabolism
    bioRxiv preprint doi: https://doi.org/10.1101/2020.07.15.204420; this version posted July 16, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Emergence of diauxie as an optimal growth strategy under resource allocation constraints in cellular metabolism Pierre Salvy1, Vassily Hatzimanikatis1,* 1 Laboratory of Computational Systems Biotechnology, Ecole´ Polytechnique F´ed´erale de Lausanne (EPFL), Lausanne, Switzerland * [email protected] Abstract The sequential rather than simultaneous consumption of carbohydrates in bacteria such as E. coli, a phenomenon termed diauxie, has been hypothesized to be an evolutionary strategy which allows the organism to maximize its instantaneous specific growth, thus giving the bacterium a competitive advantage. Currently, computational techniques used in industrial biotechnology fall short of explaining the intracellular dynamics underlying diauxic behavior, in particular at a proteome level. Some hypotheses postulate that diauxie is due to limitations in the catalytic capacity of bacterial cells. We developed a robust iterative dynamic method based on expression- and thermodynamically enabled flux models (dETFL) to simulate the temporal evolution of carbohydrate consumption and cellular growth. The dETFL method couples gene expression and metabolic networks at the genome scale, and successfully predicts the preferential uptake of glucose over lactose in E. coli cultures grown on a mixture of carbohydrates. The observed diauxic behavior in the simulated cellular states suggests that the observed diauxic behavior is supported by a switch in the content of the proteome in response to fluctuations in the availability of extracellular carbon sources.
    [Show full text]
  • Emergence of Diauxie As an Optimal Growth Strategy Under Resource Allocation Constraints in Cellular Metabolism
    Emergence of diauxie as an optimal growth strategy under resource allocation constraints in cellular metabolism Pierre Salvya and Vassily Hatzimanikatisa,1 aLaboratory of Computational Systems Biotechnology, Ecole´ Polytechnique Fed´ erale´ de Lausanne, CH-1015 Lausanne, Switzerland Edited by Jens Nielsen, BioInnovation Institute, Copenhagen, Denmark, and approved January 15, 2021 (received for review July 23, 2020) Diauxie, or the sequential consumption of carbohydrates in bac- maximization assumptions, FBA models predict the simultane- teria such as Escherichia coli, has been hypothesized to be an ous consumption of two or more carbon sources to achieve evolutionary strategy which allows the organism to maximize its the maximum possible growth (3). However, this contradicts instantaneous specific growth—giving the bacterium a competi- Monod’s observation of distinct, sequential phases of carbon tive advantage. Currently, the computational techniques used in consumption and suggests that diauxie does not come from industrial biotechnology fall short of explaining the intracellular stoichiometric constraints. dynamics underlying diauxic behavior. In particular, the under- To account for diauxie beyond stoichiometric modeling, we standing of the proteome dynamics in diauxie can be improved. looked into other biological features. Because a cell has a We developed a robust iterative dynamic method based on physiological constraint on the total amount of enzymes it can expression- and thermodynamically enabled flux models to sim- house, which we will call a proteome allocation constraint, it is ulate the kinetic evolution of carbohydrate consumption and likely that the cell will preferentially distribute its limited cat- cellular growth. With minimal modeling assumptions, we cou- alytic capacity toward pathways that utilize the most efficient ple kinetic uptakes, gene expression, and metabolic networks, substrate/enzyme combination (2, 10, 11).
    [Show full text]
  • Adenosine 3':5'-Cyclic Monophosphate As a Regulator of Bacterial Transformation (Hemophilus Influenzae Rd/Cyclic-AMP Assay) EDMUND M
    Proc. Nat. Acad. Sci. USA Vol. 70, No. 2, pp. 471-474, February 1973 Adenosine 3':5'-Cyclic Monophosphate as a Regulator of Bacterial Transformation (Hemophilus influenzae Rd/cyclic-AMP assay) EDMUND M. WISE, JR., SUSAN P. ALEXANDER, AND MARILYN POWERS Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111 Communicated by Jack L. Strominger, December 14, 1972 ABSTRACT Adenosine 3': 5'-cyclic monophosphate from overnight cultures made alkaline and then neutralized as added to exponentially growing cells of Hemophilus in- described by Goodgal and Herriott (24). Purified DNA was fluenzae strain Rd increases competence for transforma- tion 100- to 10,000-fold. Cyclic AMP added to near-station- made essentially after the procedure of Marmur (26). DNA ary or stationary cells does not increase competence over concentrations were measured by chemical (27) or spectro- the high level normally found in the early stationary scopic (28) methods. phase. Transformation Assay. Slant cultures were inoculated into In most bacteria that can take up naked DNA and can in- supplemented BHI broth and were shaken overnight at 37°. corporate genes of this DNA into its own chromosome, i.e., be In the morning 1% inocula were added to fresh broth and transformed, highest levels of competence for transformation shaken at 37°. Unless otherwise noted, neutralized 1 mM usually occur in late exponential phase and may persist into cAMP was added when the cells were at relative O.D. 0.05 at the stationary phase (1-3). Late exponential or stationary 600 nm on a Coleman Junior II Spectrophotometer (about phase is also the time when highest concentrations of internal true O.D.
    [Show full text]
  • Coordination of Bacterial Proteome with Metabolism by Cyclic AMP Signalling
    ARTICLE doi:10.1038/nature12446 Coordination of bacterial proteome with metabolism by cyclic AMP signalling Conghui You1,2, Hiroyuki Okano1,2*, Sheng Hui1,3*, Zhongge Zhang2, Minsu Kim1, Carl W. Gunderson2, Yi-Ping Wang4, Peter Lenz5, Dalai Yan6 & Terence Hwa1,2,3 The cyclic AMP (cAMP)-dependent catabolite repression effect in Escherichia coli is among the most intensely studied regulatory processes in biology. However, the physiological function(s) of cAMP signalling and its molecular triggers remain elusive. Here we use a quantitative physiological approach to show that cAMP signalling tightly coordinates the expression of catabolic proteins with biosynthetic and ribosomal proteins, in accordance with the cellular metabolic needs during exponential growth. The expression of carbon catabolic genes increased linearly with decreasing growth rates upon limitation of carbon influx, but decreased linearly with decreasing growth rate upon limitation of nitrogen or sulphur influx. In contrast, the expression of biosynthetic genes showed the opposite linear growth-rate dependence as the catabolic genes. A coarse-grained mathematical model provides a quantitative framework for understanding and predicting gene expression responses to catabolic and anabolic limitations. A scheme of integral feedback control featuring the inhibition of cAMP signalling by metabolic precursors is proposed and validated. These results reveal a key physiological role of cAMP- dependent catabolite repression: to ensure that proteomic resources are spent on distinct metabolic sectors as needed in different nutrient environments. Our findings underscore the power of quantitative physiology in unravelling the underly- ing functions of complex molecular signalling networks. Biological organisms use a myriad of signalling pathways to monitor of cAMP signalling—not for CCR per se as we will show, but for the the environment and adjust their genetic programs in accordance coordination of metabolism with proteomic resource allocation.
    [Show full text]
  • Igem 2008 Jamboree Program
    iGEM 2008 Jamboree Program Project Logi-col[i]: Terminator/Attenuator anti-sense Logic (T/AasL) Alberta NINT, Foundational Advance Rm D463, 4:00 PM Two major hurdles facing the development of complex genetic logic circuits are device connectibility and device extensibility. Connectibility refers to the ability to connect the output of one device to the input of another device, while extensibility refers to the dual abilities to rationally design new devices and combine multiple devices in one organism. Our project uses terminator/attenuator (T/A) hairpin sequences (gates) to control transcription and anti-sense RNA as input/output signals to/from the devices. We call this approach Terminator/Attenuator anti-sense Logic (T/AasL – pronounced “taw-ssel”). It solves the connectibility problems of common protein-based approaches because the anti-sense output of one device is used to disrupt formation of T/A hairpin structures of downstream devices, thus activating them. In addition, because RNA secondary structures can be rationally designed (using our m-fold derived analysis program) we can readily construct a large family of devices with minimal cross-talk for inclusion in a single cell. Differentiation and Targeting of Stem Cells to Infarcted Cardiac Tissue Bay Area RSI, Health or Medicine Rm 34-101, 11:00 AM Every year over 1.2 million people suffer myocardial infarction. The resulting heart damage requires new approaches for effective repair. Stem cell therapies provide hope. However none of the stem cell therapies currently in clinical trials addresses the need for efficient stem cell targeting to cardiac tissue or the need to replace efficiently dead tissue with new cardiomyocytes.
    [Show full text]
  • Automated Biopart Characterisation for Synthetic Biology
    Automated BioPart Characterisation for Synthetic Biology Christopher David Hirst Imperial College London Department of Bioengineering Centre for Synthetic Biology and Innovation Submitted for Degree of Doctor of Philosophy February 2014 Page | 0 Declaration All work and data contained within this thesis is my own except where referenced. ‘The copyright of this thesis rests with the author and is made available under a Creative Commons Attribution Non-Commercial No Derivatives licence. Researchers are free to copy, distribute or transmit the thesis on the condition that they attribute it, that they do not use it for commercial purposes and that they do not alter, transform or build upon it. For any reuse or redistribution, researchers must make clear to others the licence terms of this work’ Page | 1 Acknowledgments I would like to thank my primary supervisor Richard Kitney for the many helpful discussions and support during this project, allowing me to grow as a researcher. His tireless belief in the future for engineering biology has been an inspiration. I would particularly like to thank him for the many opportunities including conferences, events and experiences that I have benefited so much from but most importantly the opportunity to work on this project. I also greatly appreciate the guidance of my other supervisors Geoff Baldwin and Paul Freemont for many useful discussions about the technology in development and overall direction of the project. For my supervisors collectively I must also express my thanks for selecting me to compete as part of the Imperial iGEM team in 2008 and supervise the team in 2010, those experiences have taught me much about myself, the communication of science and re-enforced my decision to become a researcher.
    [Show full text]