利用与su1紧密连锁的Ac转座子构建玉米插入突变体库

以位于玉米第4号染色体短臂与su1紧密连锁的Ac转座子作为Ac供体,结合w22报道系,创建1 507个Ac远距离转座事件。利用基于Southern Blot的IPCR和基于酶切连接的Genome-Walking PCR方法,分离到其中56个独立的可遗传Ac/Ds(transposed Ac/Ds,tr-Ac/Ds)侧翼序列标签,补充了Ac/Ds转座插入突变体的数量。利用玉米B73的基因组序列,通过BLAST比对分析将trAc/Ds定位到了玉米基因组上,并分析了Ac/Ds插入位点附近DNA序列,其中51个tr-Ac/Ds插入序列为单拷贝序列,鉴定到39个trAc/Ds插入到了基因内部,12个Ac/Ds插入到基因间区域,进一步证明了Ac/Ds倾向于插入基因内部和低拷贝的基因组富集区。

Generation of Insertional Mutation Library in Maize Based on Activator Tightly-linked with su1

Using a donor line containing an Ac tightly linked with su1 in chromosome 4S, we constructed 1 507 long-distance transposons. Fifty-six heritable Ac/Ds flanking-sequence-tags were isolated by southern-based IPCR and digestion-ligation-based Genome-Walking PCR methods. Through BLAST analysis with the B73 genomic sequence, the position of these tags was mapped on the maize genome. Here we identify 51 tags were inserted into sequences with only one copy, 39 tr-Acs inserted into genic region and 12 tr-Acs inserted into the intragenic section. These results supported that Ac/Ds transpose with the bias of inserting into the genic and the low-copy sequence.