An improved particle bombardment for the generation of transgenic plants by direct immobilization of relleasable Tn5 transposases onto gold particles.
Plant Mol Biol
; 77(1-2): 117-27, 2011 Sep.
Article
en En
| MEDLINE
| ID: mdl-21643845
ABSTRACT
We have developed a modified particle bombardment method for plant transgenesis. An intein-tag and a 6×Cys-tag were successively fused to the N-terminus of a hyperactive Tn5 transposase. The modified transposase was immobilized on bare gold microscopic particles via covalent binding of a 6×Cys-tag sulfydryl groups to the gold surface. The tethered transposase can bind the transposon DNA in vitro to form the transposome in the absence of Mg²âº ions. After bombardment of the gold particles carrying the transposomes into the plant cells, the transposomes will be released from the carrier due to the activated self-cleavage function of intein-tag. Our data showed this procedure integrated foreign DNA into the plant genome with an increased transformation frequency as compared to the conventional particle bombardment method. A single copy insertion can also be obtained by decreasing of the assembled transposon DNA amount in relation to plant cell biomass.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Ingeniería Genética
/
Plantas Modificadas Genéticamente
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Transposasas
/
Oro
Idioma:
En
Revista:
Plant Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
/
BOTANICA
Año:
2011
Tipo del documento:
Article