Dietrich Charles R, Cui Feng, Packila Mark L, Li Jin, Ashlock Daniel A, Nikolau Basil J, Schnable Patrick S
Interdepartmental Plant Physiology Program, Iowa State University, Ames, Iowa 50011, USA.
Genetics. 2002 Feb;160(2):697-716. doi: 10.1093/genetics/160.2.697.
The widespread use of the maize Mutator (Mu) system to generate mutants exploits the preference of Mu transposons to insert into genic regions. However, little is known about the specificity of Mu insertions within genes. Analysis of 79 independently isolated Mu-induced alleles at the gl8 locus established that at least 75 contain Mu insertions. Analysis of the terminal inverted repeats (TIRs) of the inserted transposons defined three new Mu transposons: Mu10, Mu 11, and Mu12. A large percentage (>80%) of the insertions are located in the 5' untranslated region (UTR) of the gl8 gene. Ten positions within the 5' UTR experienced multiple independent Mu insertions. Analyses of the nucleotide composition of the 9-bp TSD and the sequences directly flanking the TSD reveals that the nucleotide composition of Mu insertion sites differs dramatically from that of random DNA. In particular, the frequencies at which C's and G's are observed at positions -2 and +2 (relative to the TSD) are substantially higher than expected. Insertion sites of 315 RescueMu insertions displayed the same nonrandom nucleotide composition observed for the gl8-Mu alleles. Hence, this study provides strong evidence for the involvement of sequences flanking the TSD in Mu insertion-site selection.
利用玉米Mutator(Mu)系统广泛产生突变体,这一过程利用了Mu转座子倾向于插入基因区域的特性。然而,关于Mu在基因内插入的特异性却知之甚少。对gl8位点的79个独立分离的Mu诱导等位基因进行分析发现,其中至少75个含有Mu插入。对插入转座子的末端反向重复序列(TIRs)进行分析,确定了三个新的Mu转座子:Mu10、Mu11和Mu12。大部分(>80%)插入位于gl8基因的5'非翻译区(UTR)。5'UTR内的十个位置经历了多次独立的Mu插入。对9bp靶位点重复序列(TSD)的核苷酸组成以及TSD直接侧翼序列的分析表明,Mu插入位点的核苷酸组成与随机DNA的核苷酸组成有显著差异。特别是,在位置-2和+2(相对于TSD)处观察到C和G的频率明显高于预期。315个RescueMu插入的插入位点显示出与gl8-Mu等位基因相同的非随机核苷酸组成。因此,本研究为TSD侧翼序列参与Mu插入位点选择提供了有力证据。