Coupland G, Plum C, Chatterjee S, Post A, Starlinger P
Institut für Genetik, Universität zu Köln, Federal Republic of Germany.
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9385-8. doi: 10.1073/pnas.86.23.9385.
Deletion derivatives of the maize transposable element Activator (Ac) were constructed in vitro and inserted into a kanamycin resistance gene. These constructions were then introduced into tobacco protoplasts derived from plants previously transformed with Ac. The ability of each deletion derivative to excise was measured by whether or not kanamycin-resistant tobacco calli were recovered. This allowed us to determine the length of DNA present at each terminus that is required to respond to the products expressed by the Ac element present in the genome. We show that around 200 base pairs (bp) are required at both ends for excision to occur at wild-type levels. When between 100 and 200 bp were retained at one of the ends, reduced frequencies of excision were detected. With less than 100 bp remaining at either end, no excision was detected. In addition, we show that although similar lengths of DNA are required at each terminus, the termini are not interchangeable. The significance of these data is discussed with respect to the protein(s) which interact(s) with the termini of Ac.
玉米转座因子激活子(Ac)的缺失衍生物在体外构建,并插入到卡那霉素抗性基因中。然后将这些构建体导入先前用Ac转化的植物所产生的烟草原生质体中。通过是否回收卡那霉素抗性烟草愈伤组织来测定每个缺失衍生物的切除能力。这使我们能够确定基因组中存在的Ac元件所表达的产物响应时每个末端所需的DNA长度。我们表明,两端都需要约200个碱基对(bp)才能以野生型水平发生切除。当一端保留100至200 bp之间时,检测到切除频率降低。两端剩余少于100 bp时,未检测到切除。此外,我们表明,尽管每个末端需要相似长度的DNA,但末端不可互换。就与Ac末端相互作用的蛋白质而言,讨论了这些数据的意义。