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在功能基因组研究中,插入诱变被广泛用于基因敲除。玉米Mutator转座子因其具有较高的转座活性常被用于构建大型玉米插入突变体库。本研究利用具有活性MuDR因子的玉米材料与优良玉米自交系Z31杂交,获得1000个M1单株,自交构建M2群体,研究MuDR因子在基因组中插入位点特性。利用优化的MuTAIL-PCR方法分离出695条MuDR插入位点侧翼序列,经初步生物信息学分析得到374条非冗余的插入位点,其中的298条序列能够被定位在玉米基因组物理图谱单个位点上。实验结果揭示了MuDR因子插入的一些特性:在10条染色体上随机分布,偏向于插入到基因序列中,并在某些功能基因中有明显插入偏好。
In functional genomics studies, insertion mutagenesis is widely used in gene knockout. The maize Mutator transposon is often used to construct large maize insert mutants because of its high transposase activity. In this study, maize material with active MuDR was crossed with Z31, an excellent maize inbred line, and 1000 M1 plants were obtained. The M2 population was selfed to study the locus insertion characteristics of MuDR in the genome. The flanking sequences of 695 MuDR insertion sites were isolated by optimized MuTAIL-PCR method. After preliminary bioinformatics analysis, 374 non-redundant insertion sites were found, of which 298 sequences could be mapped to a single position in the genome of maize genome Point. The experimental results revealed some of the features of MuDR factor insertion: randomly distributed on 10 chromosomes, favoring insertion into the gene sequence and significant insertion preference in certain functional genes.