CV Mattias Gustafsson

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CV Mattias Gustafsson

  1. 1. Nybogatan 17, 273 30 Tomelilla Sweden Tel: +46 (0)417 10373 Mobile: +46 (0)730 510373 Email: Mattias.Gustafsson@tomelilla.nu CURRICULUM VITAE Mattias Gustafsson SUMMARY I’m 43 years of age, got a Ph.D. in microbiology from Lund University, with 13 years of postdoctoral experience, including three years of international experience . I’ve got extensive experience in conducting biomedical research, mainly in microbiology/infection biology but also in tumor biology. I have thus a very broad experience of various lines of research and am technically quite versatile. As a person I’m curious, creative and innovative. I’m also persistent and have been productive in every workplace I’ve been to, something that is reflected in my list of scientific publications in the appendix of the CV. I have had both leading and supportive roles in the various projects I have worked on, that is , I can both be the supportive team player as well as the driving force in a project . I have proved inventive both in solving smaller technical problems in various projects as well a s making major discoveries. I have the quality to often see what is missin g in a project and thus to come up with solution to problem at hand. PROFESSIONAL EXPERIENCE Work for Professor Gunnar Lindahl 2012–present Postdoctoral Researcher Lund University Responsibilities  Conducting research on group A streptococci ( Streptococcus pyogenes ) and their interactions with the immune system.  Supervising the younger members of the laboratory .  Writing scientific articles .  Maintaining and genotyping of the transgenic mouse colony as well as other mouse work.  Responsible for the IT infrastructure of the group (installing computers and programs). Achievements  Successfully co-supervised a Ph.D. Student (I was not the official supervisor) that soon will graduate from the University of Copenhagen.  Successfully co-led a project where we invented and developed a superior method for the purification of the human blood protein Factor H. This protein can most likely be used clinically and a patent application has been filed for the invention. We submitted a patent application stemming from the conducted research.  As a member of the team I successfully contributed to an important piece of work where we discovered that many S. pyogenes class I M proteins bind complement Factor H exclusively to their hypervariable regions. We also discovered that contrary to general belief, that factor H did not have any effect on phagocytosis resistance and virulence. Since S. pyogenes is one of the major human pathogens the M protein
  2. 2. CURRICULUM VITAE Mattias Gustafsson a very important virulence factor, these findings probably have importance for understanding streptococcal diseases.   Published a first author article in the prestigious journal PLoS Pathogens (in vivo article 1 in the appendix).  2010–2012 Successfully maintained a transgenic mouse colony and conducted research on these. Submitted a last author manuscript on the Factor H purification method described above to a scientific journal. Postdoctoral Researcher University of Copenhagen Responsibilities  Conducting research on group A streptococci and their interactions with the host immune system.  Supervising the younger members of the laboratory.  Writing scientific articles .  Mouse work.  Responsible for the IT infrastructure of the group (installing computers and programs). Achievements   2008–2010 I co-supervised a Ph.D. Student (I was not the official super visor). I helped with ideas for his thesis project. These projects were successful (see above). I was involved in an ongoing project were we showed that the S. pyogenes M protein hypervariable reg ion avoids antibodies both by antigen variation and poor immunogenicity, something that is very important for understanding S. pyogenes pathology and vaccine development. I was drawn into the project when it was already ongoing. I solved a technical issue regarding the detection of antibodies which was very important for the rest of the project. This work was published in the prestigious journal Cell Host & Microbe (in vivo article 3 in the appendix) where I am shared first author. Postdoctoral Researcher Lund University Responsibilities  Conducting research on group A streptococci and their interactions with the host immune system.  Supervising the younger members of the laboratory.  Responsible for the IT infrastructure of the group (installing c omputers and programs). Achievements  Please see above. Page 2 of 10
  3. 3. CURRICULUM VITAE Mattias Gustafsson Work for Professor Catharina Svanborg 2007–2008 Postdoctoral Researcher/Microbiologist Region Skåne Responsibilities  Conducting and planning research on the HAMLET (human lactalbuming made lethal to tumor cells), a protein that kills tumor cells.  Conducting and planning research on urinary tract infections with Escherichia coli as model bacterium and the interactions with the immune system.  Supervising the younger members of the laboratory.  Teaching Achievements   I successfully introduced real time PCR in the laboratory. This led to in vitro articles 2, 3, 5 and 6 in the appendix. For article 3 I was also the intellectual sparring partner and suggested biological markers to measure.  Successfully supervised two exam project students (not the official supervisor).  Planned and conducted problem based learning (PBL) sessions at the Biomedicine program at the Medical Faculty at Lund Univers ity.  Tutored at PBL sessions at the courses General Pathology and Homeostasis at the Medicine program, Medical Faculty, Lund University.  2006–2007 I successfully introduced the use of siRNA technology in the laboratory. This led to important experiments that led to in vitro articles 2 and 6. For the project in article 2 I also gave advice in using transgenic cell lines. Published research on HAMLET and urinary tract infections in scientific articles (in vitro articles 2-6and in vivo article 4 in the appendix). Senior Researcher HAMLET Pharma Responsibilities  Conducting and planning research on the HAMLET (human lactalbuming made lethal to tumor cells), a protein that kills tumor cells.  Conducting and planning research on urinary tract infections with Escherichia coli as model bacterium and the interactions with the host immune system.  Supervising the younger members of the laboratory. Achievements  2004–2006 For scientific achievements, please see above. Postdoctoral Researcher Lund University Responsibilities  Conducting and planning research on urinary tract infections with Escherichia coli as model bacterium and the interactions with the host immune system.  Supervising the younger members of the laboratory.  Responsible for the introduction of a range of molecular biology techniques in Professor Svanborg´s laboratory.  Teaching Achievements  Successfully introduced a range of molecular biology techniques in Professor Svanborg´s research group. This fundamentall y changed the Page 3 of 10
  4. 4. CURRICULUM VITAE Mattias Gustafsson planning of projects and what was perceived as technically feasible in the laboratory.  Planned and conducted problem based learning (PBL) sessions at the Biomedicine program at the Medical Faculty at Lund University.  Produced scientific results that were later published in scientific journals. Work for Professor Colin Palmer 2001–2004 Postdoctoral Researcher University of Dundee Responsibilities  Conducting research on Peroxisome Proliferator Activated Receptors, PPARs.  Develop transgenic mice and cell lines for conditional expression of PPARs.  Maintaining and genotyping of transgenic mice.  Writing scientific articles .  Plan the work and supervise a research assistant. Achievements  Successfully developed transgenic mice and cell lines for conditional expression of human PPARδ and a dominant derivative. Interestingly  I discovered that the laboratory had chosen a system for conditional gene expression in mice that was not going to wor k. Together with Professor Palmer we chose another system.  I was recruited to a project in the laboratory investigating the role of PPARδ in the growth of breast and prostate cancer cell lines. I saw that data on breast cancer cell growth in transgenic cel l lines designed for conditional expression of PPARδ would be advantageous to the project. Such data was requested by a reviewer for in vitro article and led to its acceptance (in vitro article 8 in the appendix).  Successfully led a project in which I supe rvised a research assistant. This led to a publication (in vitro article 1 in the appendix).  The research for this postdoc period gave rise to five scientific articles (in vivo articles 2 and 5 and in vitro articles 1, 7 and 8 in the appendix). Of note, article 8, of which I am the second author, has been cited over 90 times. Work for Professor Claes von Wachenfeldt 1995–2000 Ph. D. student Lund University Responsibilities  Conducting research on cytochromes P450 in Bacillus subtilis .  Teaching  Responsibility for maintaining some laboratory equipment. Achievements  For the first part of my Ph.D. studies we collaborated with Professor Sonenshine´s group in Boston. We discovered a novel mechanism of resistance to the amino acid antibiotic 4-azaleucine in B. subtilis . We found that the resistance is due to a mutation in a transcriptional repressor leading to overproduction of two membrane proteins (then with unknown function) for branched-chain amino export. I was the first person in the group that had access to data from a range of mutant bacteria and I remember the excitement of working out the mechanism on my own (in vitro article 12 in the appendix) . Page 4 of 10
  5. 5. CURRICULUM VITAE Mattias Gustafsson  Worked at the University of Dundee with Professor Colin Palmer for a few months. This work led to in vitro article 11 in the appendix. I was offered a postdoc position and later went to work for Professor Palmer.  In the work leading to in vitro article 11 was based on discoveries in the related bacterium Bacillus megaterium , where the gene for a cytochrome P450 enzyme is preceded by the gene for a transcriptional repressor. The repressor is displaced from the DNA by certain fatty acids and the action of the P450 enzyme on the fatty acids ren der these inactive for binding the repressor. I discovered that the cytochrome P450 enzyme that we studied might have endogenous ligands that when metabolized didn’t displace the repressor from the DNA.  I initiated and led the project where I discovered th at a substance excreted by wild-type B. subtilis that can overcome the expression of a range of genes in bacteria mutated in the gene for central transcriptional regulator Spo0A (in vitro article 10 in the appendix).  Met Professor Andrew Munro at a confer ence and initiated collaboration with him and Professor Stephen Chapman at the University of Edinburgh. I suggested that we should study the activity of the cytochrome P450 enzymes studied on the endogenous branched -chain fatty acids found in the B. subtilis cell membrane. This led to in vitro publication 9 in the appendix. I was offered a postdoc position with Professor Munro.  Supervised a project student  Taught on laboratory practicals on the courses given at the Department of Microbiology  Thesis with the title ” Bacterial Cytochromes P450: Studies on cytochrome P450 102A2 and P450 102A3 of Bacillus subtilis .  Published four scientific articles (in vitro articles 9-12 in the appendix). Of note, article 9 and 12 have been cited over 50 and 60 times, respectively. Undergraduate work for Professor Mats Hansson and Professor Lars Hederstedt 1994 Undergraduate student Lund Univer Responsibilities  Conducting research on heme biosynthesis in Bacillus subtilis . Achievements  I constructed the expression systems for production of the heme biosynthesis enzyme HemY. Only a certain level of production of this enzyme is tolerated by the bacterium and I was wise enough to salvage the one or two mutants I got. These were later shown to be promoter down mutants allowing tolerable overproduction of HemY in B. subtilis .  In the same project I discovered that HemY is strongly associated with the bacterial membrane, at the time it was thought to only be loosely associated due to detergent contamination of laboratory equipment . This work resulted in in vitro article 13 in the appendix. Page 5 of 10
  6. 6. CURRICULUM VITAE Mattias Gustafsson EDUCATION 2000 Doctor of Philosophy, Department of Microbiology, Lund University with Professor Claes von Wachenfeldt as supervisor. Key focuses: Molecular biology and biochemistry of cytochromes P450 in the Gram positive model bacterium Bacillus subtilis . 1995 Bachelor of Science, Lund University , specializing in Microbiology. Key focuses: In my exam work I studied the biochemistry of the heme synthesis, see article 1 in the appendix. Courses: Chemistry, organic chemistry, biochemistry, genetics, molecular genetics, immunology, physiology, microbiology, geology, ecology, mathematics etc. TRAINING 2001 Animal course, University of Aberdeen, U.K. I held a personal license for working with experimental animals. 2005 University teaching course “Högskolepedagogisk grundkurs” , Medical Faculty, Lund University. 2005 Problem based learning course, Medical Faculty, Lund University. 2010 Laboratory animal science, Medical Faculty, Lund University . 2011 Researcher’s program, a course in commercializing research, Medeon, Malmö. LANGUAGES Swedish Native tounge English Fluent PERSONAL DATA Date of birth: 22 nd of September 1970 Martial status: Partner Camilla (42) Children: Hanna (13) Interests: Family life Nature and the environment. I’m on the board of the local chapter of Swedish Society for Nature Conservation (Naturskyddsföreningen) . I’m also a member of “Lunds Botaniska Förening”. I’m training judo and Brazilian jiu jitsu two to four times a week to keep in shape. Page 6 of 10
  7. 7. CURRICULUM VITAE Mattias Gustafsson APPENDIX Skills profile IN VIVO TECHNIQUES       Responsible for breeding transgenic mice Marking/biopsy of mice for genotyping Designed PCR-based genotyping assay for transgenic mice Dissecting mice Isolated RNA from various tissues Bleeding mice, both by heart punctuation and by cutting neck arteries     Intraperitoneal injection of mice For work regarding molecular biology techniques regarding mice, please see above Assisted in immunization of mice Assisted in infection of mice IN VITRO TECHNIQUES DNA and RNA techniques                PCR Cloning Restriction enzyme cleavage analysis Preparation of chromosomal DNA from mammalian cells, Bacillus subtilis , Streptococcus pyogenes and Escherichia coli Preparation of plasmid DNA with various methods Transformation of B. subtilis , E. coli and S. pyogenes Transfection of mammalian cells Southern blot Generation of high frequency of recombination (Hfr) strains of E. coli Mapping of genetic markers on the E. coli chromosome through mating with a Hfr-strain Mapping of genetic markers of the B. subtilis chromosome through threefactor crossings Mapping of genetic marker on the B. subtilis chromosome using a YAC library Fractionation of restriction enzyme cleaved chromosomal DNA by ultracentrifugation DNA library-construction DNA library screening             Generation of insertion and insertiondeletion B. subtilis and S. pyogenes as well as unmarked mutants in B. subtilis Transposon mutagenesis Site-directed mutagenesis Chemical mutagenesis Generation of random deletions using exonuclease III and mung bean nuclease Sequencing and publishing a DNA sequence (GenBank accession no. Y11043) Generating DNA constructs for non conditional gene-expression in B. subtilis , E. coli , mammalian cell-lines and transgenic mice Generating DNA constructs for conditional gene-expression in B. subtilis , E. coli , mammalian cell-lines and transgenic mice RNA extraction from B. subtilis , E. coli and mammalian cells Generation of cDNA from mammalian RNA Northern blot Real time PCR analysis of mammalian gene expression using Taqman and SYBR green chemistry Protein techniques       T7 protein over-production systems Disruption of cells using a sonicator Disruption of cells using a French press Membrane and cytoplasmic fraction preparations Phase fractionation of membrane proteins using Triton X114 Purification of histidine-tagged proteins using Ni-agarose columns      Purification of GST-tagged proteins using glutathione-sepharose columns Purification of proteins using ion exchange, gel filtration and ADPsepharose columns SDS-PAGE Western blot Determination of protein concentrations using various methods Page 7 of 10
  8. 8. CURRICULUM VITAE     Determination of cytochrome P450 concentration by spectrophotometry Immunocytochemistry Determination of Kd of Cytochrome P450s for ligands by spectrophotometry Determination Ki of the transcriptional repressor FatR for various ligands by EMSA Mattias Gustafsson      Determination of Kd for FatR using a direct binding assay with a fluorescent fatty acid Investigating enzyme reactions using spectrophotometric and spectrofluorometric methods ELISA Solid phase RIA Bacterial ligand and antibody binding assays Functional screening assays   Bacterial two-hybrid assays Investigating PPAR agonist induced transcriptional transactivation using mammalian one-hybrid systems   Using mammalian two-hybrid systems to investigate PPAR/RXR heterodimerisation and transactivation Investigating PPAR repression and agonist induced activation Bioinformatics      Using the GCG and Staden packages for handling smaller sequencing projects Using the GCG package for predicting open reading frames Experience with the GCG, MacVector, Vector NTI and Emboss packages Using BLAST for finding proteins similar to unknown open reading frames Using the GCG package for predicting transcriptional terminator structures     In silico design of plasmids Generating phylogenetic trees using the GCG program package Predicting transmembrane helices using the GCG package Using RasMol for looking at 3D structures of proteins Other techniques in microbiology     Gram staining Typing of bacteria using biochemical and morphological criteria Determining generation times of bacteria One step growth curves of phage T4 of E.   Titration of bacteriophages Investigating antibiotic resistance of bacteria  Phase contrast microscopy coli Microscopy   General light microscopy Fluorescence microscopy TEACHING    Taught in the laboratory practicals of the following courses given in the biology program at Lund University (three times for each course): five and ten credits Microbiology and ten credits procaryotic molecular genetics. Organized a week of the Molecular Medicine course of the Biomedicine program at Lund University for two years. Tutored and wrote problem based learning cases for one-two weeks on the Molecular Medicine course of the    Biomedicine program at Lund University (four times). Tutored for four weeks on the General Pathology course of the Medicine program, Lund University) Tutored for a half semester on the Homeostasis course of the Medicine program, Lund University. Tutored for two weeks on the Pathobiology and Pharmacology course on the Biomedicine program, Lund University. Page 8 of 10
  9. 9. CURRICULUM VITAE Mattias Gustafsson MANAGEMENT/SUPERVISION  As a Ph. D. student I supervised a project student.  As a postdoc in Dundee I led and planned the work of a research assistant . One of my main tasks in Professor Svanborg´s laboratory was to supervise members of the laboratory in molecular biology techniques .    During my appointment as postdoc with Professor Svanborg’s group I did to some extent supervise a Ph.D. student (I was not a formal supervisor). I also supervised two half semester project students (not a formal supervisor). In my current appointment I have to a large extent supervised a Ph. D. student at Copenhagen University with Professor Lindahl and Prof. John Olsen as the formal supervisors. IT SYSTEMS    Responsible for installing computers and programs in our research group. Been working with Microsoft Windows (XP, Vista and 7), Mac OS X Mountain Lion. Microsoft Office (Word, Excel, Power Point).    Experience with the GCG and Emboss packages. Experience with the Staden package. In silico cloning with MacVector and VectorNTI SCIENTIFIC PUBLICATIONS Articles In vivo 1. 2. Gustafsson, M. C., Lannergård, J., Nilsson, O. R., Kristensen, B. M., Olsen, J. E., Harris, C. L., Ufret-Vincenty, R. L., Stålhammar-Carlemalm, M., and Lindahl, G. (2013) Factor H binds to the hypervariable region of many Streptococcus pyogenes M proteins but does not promote phagocytosis resistance or acute virulence, PLoS Pathog 9, e1003323. Higgins, L. G., Garbacz, W. G., *Gustafsson, M. C., Nainamalai, S., Ashby, P. R., Wolf, C. R., and Palmer, C. N. (2012) Conditional expression of human PPARδ and a dominant negative variant of hPPARδ in vivo, PPAR research 2012, 216817. 3. Lannergård, J., Gustafsson, M. C., Waldemarsson, J., Norrby-Teglund, A., StålhammarCarlemalm, M., and Lindahl, G. (2011) The hypervariable region of Streptococcus pyogenes M protein escapes antibody attack by antigenic variation and weak immunogenicity, Cell Host & Microbe 10, 147-157. 4. Ragnarsdóttir, B., Jönsson, K., Urbano, A., Grönbe rg-Hernandez, J., Lutay, N., Tammi, M., Gustafsson, M., Lundstedt, A. C., Leijonhufvud, I., Karpman, D., Wullt, B., Truedsson, L., Jodal, U., Andersson, B., and Svanborg, C. (2010) Toll -like receptor 4 promoter polymorphisms: common TLR4 variants may prote ct against severe urinary tract infection, PLoS One 5, e10734. 5. Romanowska, M., Reilly, L., Palmer, C. N., Gustafsson, M. C., and Foerster, J. (2010) Activation of PPARβ/δ causes a psoriasis -like skin disease in vivo, PLoS One 5, e9701. *Shared first authorship in article 3. Please note that I didn’t perform any of the human in vivo experiments in article 5. In vitro/analysis of clinical samples 1. Nilsson, O. R., Lannergård, J., Morgan, B. P., Lindahl, G. and Gustafsson, M. C. (2013) Affinity purification of human factor H on polypeptides derived from streptococcal M protein: enrichment of the Y402 variant, PLoS One , Accepted for publication 2. Gustafsson, M. C., Knight, D., and Palmer, C. N. (2009) Ligand modulated antagonism of PPARγ by genomic and non-genomic actions of PPARδ, PLoS One 4, e7046. 3. Aits, S., Gustafsson, L., Hallgren, O., Brest, P., Gustafsson, M., Trulsson, M., Mossberg, A. K., Simon, H. U., Mograbi, B., and Svanborg, C. (2009) HAMLET (human alpha -lactalbumin made lethal to tumor cells) triggers autophagic tumor cell death, Int J Cancer 124, 1008-1019. 4. Brest, P., Gustafsson, M., Mossberg, A. K., Gustafsson, L., Düringer, C., Hamiche, A., and Svanborg, C. (2007) Histone deacetylase inhibitors promote the tumoricidal effect of HAMLET, Cancer Res 67, 11327-11334. Page 9 of 10
  10. 10. CURRICULUM VITAE Mattias Gustafsson 5. Lundstedt, A. C., McCarthy, S., Gustafsson, M. C., Godaly, G., Jodal, U., Karpman, D., Leijonhufvud, I., Lindén, C., Martinell, J., Ragnarsdóttir, B., Samuelsson, M., Truedsson, L., Andersson, B., and Svanborg, C. (2007) A genetic basis of susceptibility to acute pyelonephritis, PLoS One 2, e825. 6. Ragnarsdóttir, B., Samuelsson, M., Gustafsson, M. C., Leijonhufvud, I., Karpman, D., and Svanborg, C. (2007) Reduced toll-like receptor 4 expression in children with asymptomatic bacteriuria, J Infect Dis 196, 475-484. 7. Fischer, H., Ellström, P., Ekström, K., Gustafsson, L., Gustafsson, M., and Svanborg, C. (2007) Ceramide as a TLR4 agonist; a putative signalling intermediate between sphingolipid receptors for microbial ligands and TLR4, Cell Microbiol 9, 1239-1251. 8. Targett-Adams, P., McElwee, M. J., Ehrenborg, E., Gustafsson, M. C., Palmer, C. N., and McLauchlan, J. (2005) A PPAR response element regulates transcription of the gene for human adipose differentiation-related protein, Biochim Biophys Acta 1728, 95-104. 9. Stephen, R. L., Gustafsson, M. C., Jarvis, M., Tatoud, R., Marshall, B. R., Knight, D., Ehrenborg, E., Harris, A. L., Wolf, C. R., and Palmer, C. N. (2004) Activation of peroxisome proliferator-activated receptor δ stimulates the proliferation of human breast and prostate cancer cell lines, Cancer Res 64, 3162-3170. 10. Gustafsson, M. C., Roitel, O., Marshall, K. R., Noble, M. A., Chapman, S. K., Pessegueiro, A., Fulco, A. J., Cheesman, M. R., von Wachenfeldt, C., and Munro, A. W. (2004) Expression, purification, and characterization of Bacillus subtilis cytochromes P450 CYP102A2 and CYP102A3: flavocytochrome homologues of P450 BM3 from Bacillus megaterium , Biochemistry 43, 5474-5487. 11. Gustafsson, M. C., and von Wachenfeldt, C. (2001) A novel diffusible substance can overcome the apparent AbrB repression of the Bacillus subtilis fatR promoter, FEMS Microbiol Lett 199, 197-202. 12. Gustafsson, M. C., Palmer, C. N., Wolf, C. R., and von Wachenfeldt, C. (2001) Fatty -aciddisplaced transcriptional repressor, a conserved regulator of cytochrome P450 102 transcription in Bacillus species, Arch Microbiol 176, 459-464. 13. Belitsky, B. R., Gustafsson, M. C., Sonenshein, A. L., and von Wachenfeldt, C. (1997) An lrp like gene of Bacillus subtilis involved in branched-chain amino acid transport, J Bacteriol 179, 5448-5457. 14. Hansson, M., Gustafsson, M. C., Kannangara, C. G., and Hederstedt, L. (1997) Isolated Bacillus subtilis HemY has coproporphyrinogen III to coproporphyrin III oxidase activity, Biochim Biophys Acta 1340, 97-104. Reviews 1. Ragnarsdóttir, B., Fischer, H., Godaly, G., Grönberg -Hernandez, J., Gustafsson, M., Karpman, D., Lundstedt, A. C., Lutay, N., Rämisch, S., Svensson, M. L., Wullt, B., Yadav, M., and Svanborg, C. (2008) TLR- and CXCR1-dependent innate immunity: insights into the genetics of urinary tract infections, Eur J Clin Invest 38 Suppl 2, 12-20. 2. Svanborg, C., Bergsten, G., Fischer, H., Godaly, G., Gustafsson, M., Karpman, D., Lundstedt, A. C., Ragnarsdóttir, B., Svensson, M., and Wullt, B. (2006) Uropathogenic Escherichia coli as a model of host-parasite interaction, Curr Opin Microbiol 9, 33-39. 3. Palmer, C. N., Gustafsson, M. C., Dobson, H., von Wachenfeldt, C., and Wolf, C. R. (1999) Adaptive responses to fatty acids are mediated by the regulated expression of cytochromes P450, Biochem Soc Trans 27, 374-378. Ph.D. thesis 1. Gustafsson, M. C. U. (2000) Bacterial cytochromes P450: Studies on cytochromes P450 102A2 and P450 102A3 of Bacillus subtilis., Department of Microbiology, Lund University, Lund, Sweden. Page 10 of 10

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