Expression of Human Lactoferrin in Bacteroides uniformis and its Effect on Azoxymethane-induced Aberrant Crypt Focus Formation in the Rat Colon |
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| Institution: | 1. Department of Bacteriology, School of Medicine, The University of Tokushima, Kuramoto-cho, Tokushima, 770-8503, Japan;2. Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand;3. Animal Molecular Physiology Research Unit, Korea Research Institute of Bioscience and Biotechnology, Taejon, Korea;4. Experimental Pathology and Chemotherapy Division, National Cancer Center Research Institute, Tokyo, 104-0045, Japan;1. Helmholtz-Zentrum Dresden-Rossendorf, Helmholtz Institute Freiberg for Resource Technology, Halsbrücker Straße 34, 09599 Freiberg, Germany;2. Helmholtz Zentrum Dresden-Rossendorf, Institute of Resource Ecology, Bautzner Landstraße 400, 01328 Dresden, Germany;1. Institute of Biomaterials, University of Erlangen-Nuremberg, Cauerstr. 6, 91058 Erlangen, Germany;2. Department of Hand, Plastic and Reconstructive Surgery—Burn Center, BG Trauma Center Ludwigshafen/Rhine, Hand and Plastic Surgery, University Heidelberg, Ludwig-Guttmann-Str. 13, Ludwigshafen, Germany;3. Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, Switzerland;4. Clinic of Preventive Dentistry, Periodontology and Cariology, University of Zurich, Centre of Dental Medicine, Plattenstr. 11, CH-8032 Zurich, Switzerland;5. Institute for Polymer Materials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Erlangen, Germany;6. Section of Experimental Oncology and Nanomedicine, ENT-Department, University Hospital Erlangen, Germany;1. College of Polymer Science and Engineering, Sichuan University, Chengdu, Sichuan Province, 610065, PR China;2. Department of Oncology, the 452 Hospital of Chinese PLA, Chengdu, Sichuan Province, 610021, PR China;1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;2. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;3. Department of Biochemistry and Molecular Biology, Monash University, Clayton VIC3800, Australia;1. Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, PR China;2. National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, Wuxi 214122, PR China;3. Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark;4. Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi 214122, PR China;1. College of Food Science and Engineering, Tianjin University of Science & Technology, Tianjin 300457, China;2. College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China;3. College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua East Road, Haidian District, Beijing, China |
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| Abstract: | To express a human lactoferrin (hLF) gene in Bacteroides uniformis, a member of the anaerobic microflora in the colon, we constructed a recombinant plasmid, pVLFK, by subcloning hLF cDNA to an Escherichia coli–Bacteroides shuttle vector, pVAL-1. The plasmid pVLFKwas transferred to B. uniformis strain BU1001 by the filter mating procedure creating strain TCTK101. The lactoferrin protein in B. uniformis strain TCTK101 was detected by Western blot analysis with an anti-human lactoferrin monoclonal antibody. A culture of strain TCTK101 inhibited the growth ofE. coli strain HB101 in vitro compared to a culture of strain TCTK11, which is a B. uniformis strain-carrying plasmid pVAL-1, suggesting that the lactoferrin produced from strain TCTK101 possesses biological activity. To determine the effect of lactoferrin-producing B. uniformis on the formation of azoxymethane-induced aberrant crypt foci (ACF), putative neoplastic lesions, overnight cultures of strains TCTK11 and TCTK101 were given to rats as drinking water. The numbers of ACF and ACF having more than three crypts per focus five weeks after the beginning of the experiment significantly increased in the group treated by a culture of strain TCTK11 compared with those in the non-treated water group. However, rats treated with a culture of strain TCTK101 carrying plasmid pVLFK showed a significantly lower number of ACF than rats with a culture of strain TCTK11 (34% reduction), suggesting that hLF which is produced in a prokaryotic expression system prevents formation of ACF in the rat colon. |
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