Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – Involving particle-mediated transfecion
Patent
1998-06-05
2000-10-31
Fox, David T.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
Involving particle-mediated transfecion
800278, 435420, 435424, 435430, 435431, 435470, A01H 400, A01H 100, C12N 504, C12N 1582, C12N 1587
Patent
active
061405558
ABSTRACT:
Maize tissue may be regenerated from nodal extracts prepared from germinated mature seeds and germinated embryos. Nodal section explants are secured from seedlings in 3-5 days. The explants are grown on an induction medium until adventitious shoot formation is observed. The shoots are separated and elongated on an MS-based medium, and then rooted. Fast genotype-independent regeneration is obtained, in 12-14 weeks. These explants, as well as zygotic embryos, may be transformed with exogenous DNA using a biolistic approach, where DNA precipitated onto tungsten microprojectiles is accelerated as 650 psi towards the explants at a distance of at least 7.5 microns. Improved frequency of transformation is obtained using microprojectiles which prior to DNA precipitation were frozen in glycerol, and suspending from a preparation of 2.5 M CaCl.sub.2. The combination of transformation process and regeneration can be used, independent of genotype, to provide new commercial crop organisms.
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Reichert Nancy A.
Rudraswamy Vanishree
Fox David T.
Grunberg Anne Marie
Kelber Steven B.
Mississippi State University
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