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Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi.

Publication ,  Journal Article
Armaleo, D; Ye, GN; Klein, TM; Shark, KB; Sanford, JC; Johnston, SA
Published in: Current genetics
February 1990

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 micron 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.

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Published In

Current genetics

DOI

EISSN

1432-0983

ISSN

0172-8083

Publication Date

February 1990

Volume

17

Issue

2

Start / End Page

97 / 103

Related Subject Headings

  • Transformation, Genetic
  • Saccharomyces cerevisiae
  • Saccharomyces
  • Neurospora crassa
  • Neurospora
  • Microbiology
  • DNA, Fungal
  • Culture Media
  • Cell Nucleus
  • 3107 Microbiology
 

Citation

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Armaleo, D., Ye, G. N., Klein, T. M., Shark, K. B., Sanford, J. C., & Johnston, S. A. (1990). Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi. Current Genetics, 17(2), 97–103. https://doi.org/10.1007/bf00312852
Armaleo, D., G. N. Ye, T. M. Klein, K. B. Shark, J. C. Sanford, and S. A. Johnston. “Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi.Current Genetics 17, no. 2 (February 1990): 97–103. https://doi.org/10.1007/bf00312852.
Armaleo D, Ye GN, Klein TM, Shark KB, Sanford JC, Johnston SA. Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi. Current genetics. 1990 Feb;17(2):97–103.
Armaleo, D., et al. “Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi.Current Genetics, vol. 17, no. 2, Feb. 1990, pp. 97–103. Epmc, doi:10.1007/bf00312852.
Armaleo D, Ye GN, Klein TM, Shark KB, Sanford JC, Johnston SA. Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi. Current genetics. 1990 Feb;17(2):97–103.
Journal cover image

Published In

Current genetics

DOI

EISSN

1432-0983

ISSN

0172-8083

Publication Date

February 1990

Volume

17

Issue

2

Start / End Page

97 / 103

Related Subject Headings

  • Transformation, Genetic
  • Saccharomyces cerevisiae
  • Saccharomyces
  • Neurospora crassa
  • Neurospora
  • Microbiology
  • DNA, Fungal
  • Culture Media
  • Cell Nucleus
  • 3107 Microbiology