Gosselé V, van Aarssen R, Cornelissen M
Plant Genetic Systems NV, Gent, Belgium.
Plant Mol Biol. 1994 Dec;26(6):2009-12. doi: 10.1007/BF00019512.
6' gentamicin acetyltransferases detoxify aminoglycoside antibiotics containing a 6' amino group. We tested whether a 6' gentamicin acetyltransferase gene (6' gat) of Shigella sp. is suitable as selectable gene in plant transformation using kanamycin (Km) as a substrate. A comparative transformation experiment using Nicotiana tabacum SR1 protoplasts showed that 6' gat is as effective for selection of transformants as the commonly used neomycin phosphotransferase II (nptII). In stably transformed plants we detected moderate levels of the 6' gat mRNA. An enzymatic assay was developed with which the acetyltransferase activity of the protein is easily demonstrated.
6′-庆大霉素乙酰转移酶可使含有6′-氨基的氨基糖苷类抗生素解毒。我们测试了志贺氏菌属的一个6′-庆大霉素乙酰转移酶基因(6′-gat)是否适合作为植物转化中的选择基因,使用卡那霉素(Km)作为底物。一项使用烟草SR1原生质体的比较转化实验表明,6′-gat在选择转化体方面与常用的新霉素磷酸转移酶II(nptII)一样有效。在稳定转化的植物中,我们检测到了中等水平的6′-gat mRNA。我们开发了一种酶活性测定方法,通过该方法可以轻松证明该蛋白质的乙酰转移酶活性。