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。
Insertional mutagenesis has now been widely used to knockout genes for functional genomics.The maize Mutator transposons hold an advantage of high activity to construct large mutant libraries.In this study,a MuDR line was used to cross with an elite Chinese maize inbred line Z31.A total of 1 000 M1 individuals were planted and self-pollinated to generate their M2 families.Experiments were conducted to investigate the insertion specificity of MuDR related transposable elements.Six hundred and ninety-five MuDR inserted flanking sequences were isolated with a modified MuTAIL-PCR method and analyzed with bioinformatics.Three hundred and seventy-four non-redundant insertion sites were identified and 298 of them were mapped to a single locus on the integrated maize map.The results revealed some prominent features of the MuDR-related insertions of maize:random distribution across the 10 chromosomes,preferential insertion into genic sequence and favoring some classes of functional genes.