BOT 4734C Chapter Notes - Chapter 3: Tungsten, Chlamydomonas, Chloroplast
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Allen (cornell university) in 1984: coi(cid:374)ed the ter(cid:373) (cid:862)biolisti(cid:272)(cid:863) whi(cid:272)h is a(cid:374) a(cid:272)ro(cid:374)y(cid:373) for (cid:271)iologi(cid:272)al (cid:271)allisti(cid:272)s, fairly simple concept. Employs high velocity metal particles (micro projectiles) to deliver reagents into cells: versatility. Any in any organelles (nucleus, plastids, and mitochondrial genomes) Landmark demonstrations: dna and rna into onion (klein et al. 1987, nature 327, 70-73: transformed chloroplast genome of chlamydomonas (boynton et al. Science 240, 1534-1538: transform mitochondrial genome of yeast (johnston et al. Particle bombardment: uses tungsten or gold particles, 0. 6~1 micrometers in diameter, coated with the dna that you want to insert into the genome, particle coating. Then ethanol washes and air dry: dna-coated particles are propelled by high pressure gas (usually helium) into the cells. Usually at a pressure of 900-1300 psi. Expression of transgenes: other materials needed: Laminar flow hood (to prevent any potential contamination of the product)
