Transformation ofSchizophyllum commune: An analysis of specific properties |
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| Institution: | 1. Department of Botany, University of Vermont, Burlington, Vermont 05405 USA;2. Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, Vermont 05405 USA;1. Department of Anatomy, Vishwabharati Medical College, Kurnool, Andhra Pradesh, India;2. Department of Anatomy, Kasturba Medical College, Manipal University, Manipal 576104, India;1. Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland;5. Metabolic Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland;2. Computational and Statistical Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland;4. Genomics Core/Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland;7. Cancer Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland;3. Nuclear Medicine Division, Clinical Center, National Institutes of Health, Bethesda, Maryland;6. Radiology and Imaging Sciences, Clinical Center, National Institutes of Health, Bethesda, Maryland;8. Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland;11. Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland;9. Information Engineering Branch, National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, Maryland;10. Computational Biology Branch, National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, Maryland;1. Institute of Chemical Materials, China Academy of Engineering Physics, Mianyang, 621900, China;2. Institute of Nuclear Physics and Chemistry, China Academy of Engineering Physics, Mianyang, 621000, China;3. Division of Nanomaterials and Chemistry Hefei National Laboratory for Physical Sciences at Microscale, University of Science and Technology of China, China;4. College of Environment and Safety Engineering, North University of China, Taiyuan, 030051, PR China;1. State Key Program of Microbiology and Department of Microbiology, College of Life Sciences, Nankai University, Tianjin 3 00071, China;2. Institute of Biochemistry and Molecular Biology, College of Life Sciences, Beijing Normal University (CLS-BNU), Beijing 100875, China;1. National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899, USA;2. Maryland State Police Forensic Sciences Division, 221 Milford Mill Road, Pikesville, MD 21208, USA;3. Vermont Forensic Laboratory, 45 State Drive, Waterbury, VT 05676, USA;1. Department of Civil and Environmental Engineering, Case Western Reserve University, 2104 Adelbert Road, Bingham 249C, Cleveland, OH, 44106, USA;2. Department of Civil and Environmental Engineering, Case Western Reserve University, 2104 Adelbert Road, Bingham 248, Cleveland, OH, 44106, USA;3. Shanghai Key Laboratory for Digital Maintenance of Buildings and Infrastructure, Department of Civil Engineering, Shanghai Jiao Tong University, 800 Dongchuan Rd, Shanghai, 200240, China;4. Department of Mechanical and Aerospace Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106, USA;5. Department of Macromolecular Science and Engineering, Case Western Reserve University, 2100 Adelbert Rd, Cleveland, OH, 44106, USA;6. Department of Macromolecular Science and Engineering, Case Western Reserve University, Cleveland, OH, 44106, USA;7. Urban and Environmental Studies, Department of Mechanical and Aerospace Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106, USA;8. Department of Civil and Environmental Engineering, Case Western Reserve University, 2104 Adelbert Road, Bingham 237, Cleveland, OH, 44106, USA |
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| Abstract: | Several properties of transformation in the basidiomycete,Schizophyllum commune, were examined. The transformation efficiency of protoplasts made from germinating basidiospores is dependent upon the length of time that the spores are incubated under conditions that promote germination. Protoplasts prepared from ungerminated spores transform at least 10 times more efficiently than protoplasts prepared from germlings (25 μm in length) or from mycelium. Transformation frequencies of 1000 transformants/μg of control plasmid DNA and 107 protoplasts are sufficient for obtaining transformants with 2 × 107 protoplasts and 10 μg of bank DNA from a genomic plasmid library. The probability of cotransforming with two plasmids is dependent on the DNA concentrations of each; concentrations can be adjusted to yield nearly 100% cotrasformants. The presence of a nonselected plasmid in the reaction mix improves the transformation frequency of a selected marker carried on another plasmid; this is not true if linear fragments ofSchizophyllum genomic DNA are used as the nonselected DNA. Transformation of aSchizophyllum protoplast does not require its fusion to another protoplast. |
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