caa a22 4500 465758924 CHVBK 20180323111855.0 cr unu---uuuuu 170327e19900201xx s 000 0 eng 10.1007/BF00312852 doi (NATIONALLICENCE)springer-10.1007/BF00312852 Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi [Elektronische Daten] [Daniele Armaleo, Guang-Ning Ye, Theodore Klein, Katherine Shark, John Sanford, Stephen Johnston] Summary: Tungsten microprojectiles coated with nucleic acid and accelerated to velocities of approximately 500 m/s, can penetrate living cells and tissues with consequent expression of the introduced genes (Klein et al. 1987). Saccharomyces cerevisiae is used here as a model system to define the basic parameters governing the biolistic (biological-ballistic) delivery of DNA into cells. Among the physical factors affecting the efficiency of the process in yeast are the microprojectile's constitution, size, concentration and amount, and the procedure used for binding DNA to it. The biological parameters that affect the process include the cell's genotype, growth phase, plating density, and the osmotic composition of the medium during bombardment. By optimizing these physical and biological parameters, rates of transformation between 10-5 and 10-4 were achieved. Stable nuclear transformants result primarily from penetration of single particles of 0.5-0.65 μm in diameter, delivering on average 10-30 biologically active plasmids into the cell. The tungsten particles detectably increase the buoyant density of the transformants' progenitors. Springer-Verlag, 1990 Fungal transformation nationallicence Particle gun nationallicence Microprojectiles nationallicence Biolistic gene transfer nationallicence Armaleo Daniele Department of Botany, Duke University, 27706, Durham, NC, USA aut Ye Guang-Ning Department of Horticultural Sciences, Cornell University, 14456, Geneva, NY, USA aut Klein Theodore Plant Gene Expression Center, USDA-ARS, 800 Buchanan St., 94710, Albany, CA, USA aut Shark Katherine Department of Horticultural Sciences, Cornell University, 14456, Geneva, NY, USA aut Sanford John Department of Horticultural Sciences, Cornell University, 14456, Geneva, NY, USA aut Johnston Stephen Department of Botany, Duke University, 27706, Durham, NC, USA aut Current Genetics Springer-Verlag 17/2(1990-02-01), 97-103 0172-8083 17:2<97 1990 17 294 https://doi.org/10.1007/BF00312852 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00312852 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Armaleo Daniele Department of Botany, Duke University, 27706, Durham, NC, USA aut NATIONALLICENCE 700 1- Ye Guang-Ning Department of Horticultural Sciences, Cornell University, 14456, Geneva, NY, USA aut NATIONALLICENCE 700 1- Klein Theodore Plant Gene Expression Center, USDA-ARS, 800 Buchanan St., 94710, Albany, CA, USA aut NATIONALLICENCE 700 1- Shark Katherine Department of Horticultural Sciences, Cornell University, 14456, Geneva, NY, USA aut NATIONALLICENCE 700 1- Sanford John Department of Horticultural Sciences, Cornell University, 14456, Geneva, NY, USA aut NATIONALLICENCE 700 1- Johnston Stephen Department of Botany, Duke University, 27706, Durham, NC, USA aut NATIONALLICENCE 773 0- Current Genetics Springer-Verlag 17/2(1990-02-01), 97-103 0172-8083 17:2<97 1990 17 294 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer