期刊
BIOLOGIA PLANTARUM
卷 54, 期 4, 页码 777-780出版社
ACAD SCIENCES CZECH REPUBLIC, INST EXPERIMENTAL BOTANY
DOI: 10.1007/s10535-010-0141-9
关键词
Helios biolistic gun; moss protonemal tissue; particle size; random and targeted integration; regeneration
资金
- Grant Agency of the Czech Republic [521/04/0971]
- Grant Agency of AS CR [S5038304]
- Ministry of Education, Youth and Sports of the Czech Republic [1M0505, LC06004]
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 mu m gold particles yielded between 20 and 40 stable transformants per 1 mu g of DNA. Transformation with circular plasmids generates higher frequencies of random transgene integration, whereas linear plasmids are more efficient in generating gene-targeted insertions.
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