Grupo de Biotecnología de Frutales, Departamento de Mejora, CEBAS-CSIC, Campus Universitario de Espinardo, Apartado de correos 164, 30100, Murcia, Spain.
Plant Cell Rep. 2010 Nov;29(11):1203-13. doi: 10.1007/s00299-010-0900-2. Epub 2010 Jul 20.
Plant transformation protocols generally involve the use of selectable marker genes for the screening of transgenic material. The bacterial gene nptII, coding for a neomycin phosphotransferase, and the hpt gene, coding for a hygromycin phosphotransferase, are frequently used. These enzymes detoxify aminoglycoside antibiotics by phosphorylation, thereby permitting cell growth in the presence of antibiotics. Nevertheless, the screening for transgenic regenerated shoots is often partial and difficult due to regeneration of escapes and chimeras. These difficulties can be caused, in part, by an incorrect assumption about the mode of action of antibiotics in bacterial and eukaryotic cells and in in vitro tissue culture. The information contained in this review could be useful to establish better selection strategies by taking into account factors such as explant complexity, transformation and selection protocols that allow better accessibility to cells of Agrobacterium and antibiotics, and faster regeneration methods that avoid collateral effects of antibiotics on recovered, putative transgenic shoots.
植物转化方案通常涉及使用选择标记基因来筛选转基因材料。细菌基因 nptII 编码新霉素磷酸转移酶，hpt 基因编码潮霉素磷酸转移酶，它们经常被使用。这些酶通过磷酸化使氨基糖苷类抗生素失活，从而允许细胞在抗生素存在的情况下生长。然而，由于逃逸和嵌合体的再生，对转基因再生芽的筛选通常是不完全和困难的。这些困难部分是由于对抗生素在细菌和真核细胞以及体外组织培养中的作用模式的不正确假设造成的。本综述中包含的信息可以通过考虑以下因素来建立更好的选择策略，这些因素包括外植体的复杂性、转化和选择方案，这些方案可以更好地使农杆菌和抗生素进入细胞，并采用更快的再生方法，避免抗生素对回收的、假定的转基因芽的间接影响。
