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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。
A Ac transposon located on the short arm of maize chromosome 4 closely linked to su1 was used as an Ac donor. In combination with the w22 reporter system, 1 507 Ac distant transposition events were established. Fifty-six independent transposable Ac / Ds (tr-Ac / Ds) flanking sequences were isolated by Southern Blot-based IPCR and restriction enzyme-based Genome-Walking PCR. Ds transposable insert the number of mutants. Using the genome sequence of maize B73, the trAc / Ds was mapped to the maize genome by BLAST analysis and the DNA sequence near the Ac / Ds insertion site was analyzed. The 51 tr-Ac / Ds insertions were single copy sequences, It was identified that 39 trAc / Ds were inserted into the gene and 12 Ac / Ds were inserted into the intergenic region, further demonstrating that Ac / Ds tend to insert into the gene interior and low-copy genomic enrichment regions.