Belzile F, Yoder J I
Department of Vegetable Crops, University of California, Davis 95616-8746.
Plant J. 1992 Mar;2(2):173-9.
A transgenic tomato line containing between eight and ten copies per genome of an exceptionally active maize transposable element Ac has previously been described. Southern analyses indicated that these elements are somatically active in these plants. In order to characterize further the pattern of somatic transposition in this line, 24 independent Ac insertion events from a single plant were cloned. In 21 cases, Ac inserted into single copy genomic DNA while in three cases Ac inserted into sequences present at two to four copies per genome; none of the insertions occurred into more highly repetitive DNA. The chromosomal locations of 20 insertion sites were determined by RFLP mapping and a pattern of small dispersed clusters emerged. Thirteen of the 20 insertion sites were linked to at least one other insertion site but these were distributed over nine of the 12 tomato chromosomes. Only one Ac insertion was linked to the T-DNA locus. The structural integrity of these Ac elements was examined and no evidence of deletions or other rearrangements suggestive of Ds elements was found. The implications of these findings with respect to the use of Ac as a transposon tag in heterologous species are discussed.
先前已描述了一种转基因番茄品系,其每个基因组含有8到10个拷贝的异常活跃的玉米转座元件Ac。Southern分析表明这些元件在这些植物中是体细胞活跃的。为了进一步表征该品系中体细胞转座的模式,从一株植物中克隆了24个独立的Ac插入事件。在21个案例中,Ac插入到单拷贝基因组DNA中,而在3个案例中,Ac插入到每个基因组有2到4个拷贝的序列中;没有插入发生在更高重复的DNA中。通过RFLP作图确定了20个插入位点的染色体位置,出现了小的分散簇模式。20个插入位点中的13个与至少一个其他插入位点连锁,但它们分布在12条番茄染色体中的9条上。只有一个Ac插入与T-DNA位点连锁。检查了这些Ac元件的结构完整性,未发现缺失或其他提示Ds元件的重排证据。讨论了这些发现对于在异源物种中使用Ac作为转座子标签的意义。