Selectable/reporter gene for use during genetic engineering of p

Chemistry: molecular biology and microbiology – Spore forming or isolating process

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435 691, 4353201, 4351723, 4352523, A01H 104, C12N 514, C12N 1500

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054749298

ABSTRACT:
Adenosine deaminase gene (ADA gene) is used as a selectable/reporter gene for plant cells and plants. When introduced into plant cells, the ADA gene makes the cells resistant to inhibition by 2-deoxyadenosine or an analog, so transformed cells can be separated from non-transformed cells by culturing a cell mixture in a medium containing 2-deoxyadenosine or an analog in amounts that inhibit normal plant cells. A single vector may be constructed containing an ADA gene from any source (e.g. a mouse ADA gene) and an additional gene-of-interest to be introduced into plant cells. The vector may then be introduced into the cells by such a route as Agrobacterium infection or micro-projectile bombardment. The presence of the ADA gene in successfully transformed plant cells may be confirmed by cultivating the cells in a medium containing 2-deoxyadenosine or an analog in the presence of a material that changes color in the presence of ammonia. Observation of a color change confirms the presence of the gene. DNA constructs containing both ADA genes and foreign genes-of-interest may be constructed, e.g. plasmid pRD360-ADA (ATCC 69459).

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