Pogorelko Gennady V, Fursova Oksana V
Nikolai Ivanovich Vavilov Institute of General Genetics, Russian Academy of Sciences, Gubkin Street 3, 119991 Moscow, Russia.
J Genet. 2008 Aug;87(2):133-40. doi: 10.1007/s12041-008-0020-8.
The exact localization of an insertion in the genome of transgenic plants obtained by Agrobacterium-mediated transformation is an integral part of most experiments aimed at studying these types of mutants. There are several methods for isolating unknown nucleotide sequences of genomic DNA which flank the borders of T-DNA integrated in the genome of plants. However, all the methods based on PCR have limitations which in some cases do not permit the desired objective to be achieved. We have developed a new technique for isolating flanking sequence tags (FSTs) via modified inverse PCR. This method is highly efficient and simple, but also retains the advantages of previously well-documented approaches.
通过农杆菌介导转化获得的转基因植物基因组中插入片段的确切定位，是大多数旨在研究这类突变体的实验不可或缺的一部分。有几种方法可用于分离整合在植物基因组中的T-DNA边界两侧的基因组DNA未知核苷酸序列。然而，所有基于PCR的方法都有局限性，在某些情况下无法实现预期目标。我们开发了一种通过改良反向PCR分离侧翼序列标签（FST）的新技术。该方法高效且简单，同时还保留了先前充分记录的方法的优点。
