Čeština
EnglishEfficient biolistic transformation of the moss Physcomitrella patens
Šmídková M., Holá M., Angelis K.J.
BIOLOGIA PLANTARUM 54: 777-710, 2010
Klíčová slova: Helios biolistic gun, moss protonemal tissue, particle size, random and targeted integration, regeneration
Abstrakt: High rates of homologous recombination (HR) in comparison to other plants make the moss Physcomitrella patens an attractive model organism for genetic studies as well as biotechnological applications. We describe a simple protocol for the efficient biolistic transformation of protonemal tissue with minimum tissue handling steps. The transformation efficiency depends on the biolistic conditions. The bombardment of tissue with 1 μm gold particles yielded between 20 and 40 stable transformants per 1 μg of DNA. Transformation with circular plasmids generates higher frequencies of random transgene integration, whereas linear plasmids are more efficient in generating gene-targeted insertions.
DOI:
Autoři z ÚEB: Karel J. Angelis
BIOLOGIA PLANTARUM 54: 777-710, 2010
Klíčová slova: Helios biolistic gun, moss protonemal tissue, particle size, random and targeted integration, regeneration
Abstrakt: High rates of homologous recombination (HR) in comparison to other plants make the moss Physcomitrella patens an attractive model organism for genetic studies as well as biotechnological applications. We describe a simple protocol for the efficient biolistic transformation of protonemal tissue with minimum tissue handling steps. The transformation efficiency depends on the biolistic conditions. The bombardment of tissue with 1 μm gold particles yielded between 20 and 40 stable transformants per 1 μg of DNA. Transformation with circular plasmids generates higher frequencies of random transgene integration, whereas linear plasmids are more efficient in generating gene-targeted insertions.
DOI: