Departamento de Fisiología Vegetal, Facultad de Biología, Campus Vida, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Galicia, Spain.
J Biotechnol. 2013 Feb 20;163(4):386-90. doi: 10.1016/j.jbiotec.2012.11.014. Epub 2012 Dec 5.
The use of minimal cassettes (linear DNA comprising promoter+open reading frame+terminator) for genetic transformation offers a means of transforming plant cells by biolistics without introducing unwanted sequences. However, a species-specific approach of the factors involved in successful transformation with this technology is advisable. Protection of the minimal cassette upstream promoter and downstream terminator may be necessary due to the nuclease activity of target plant material. Genetic transformation by biolistics followed by expression analysis was used to evaluate different DNA constructs in 110 Richter (Vitis sp.) cell suspensions. Our results suggest the importance of 3'-end cassette protection for successful protein expression using the minimal cassette technology.
使用最小的基因盒(由启动子+开放阅读框+终止子组成的线性 DNA)进行遗传转化为利用生物弹粒转化植物细胞提供了一种方法,而不会引入不需要的序列。然而,对于这种技术的成功转化所涉及的因素,采用一种物种特异性的方法是明智的。由于靶植物材料的核酸酶活性,可能需要保护最小基因盒上游启动子和下游终止子。通过生物弹粒进行遗传转化,然后进行表达分析,用于评估 110 里希特(Vitis sp.)细胞悬浮液中的不同 DNA 构建体。我们的结果表明,在使用最小基因盒技术进行成功的蛋白质表达时,保护 3'-末端基因盒的重要性。