Mutator转座子介导的玉米甜质突变体侧翼序列克隆

Cloning Flanking Sequences Sweet Corn Mutant of Maize Inserted by Mutator Transposon

为了探讨Mu转座子使玉米发生甜质突变的分子机理,利用Mu-AFLP方法分离Mutator转座子插入位点的侧翼序列,并根据侧翼序列的延伸设计一对特异性引物P1、P2,验证转座子插入的真实性,同时对插入位点所在的基因进行生物信息学分析。结果显示:侧翼序列长299 bp,插入位点位于第3染色体,该转座子的插入属于真实插入;突变基因全长5 746 bp,共编码592个氨基酸;所编码蛋白的理论分子量为67.5 k Da,疏水性氨基酸含量为42.06%,具有10个跨膜结构域,属于亲水性内膜蛋白。该结果为更好地利用Mutator转座子创制甜玉米新种质奠定了基础,也对揭示玉米胚乳发育与淀粉合成机制具有重要意义。

In order to investigate the molecular mechanism of Mu gene transfer in sweet corn mutant,the flanking sequence of the insertion site of Mutator gene was isolated by Mu-AFLP method,and the authenticity of transposon insertions was verified by a pair of specific primers P1 and P2 designed according to the extension of the flanking sequences. Meanwhile the genetic biological information of the insertion site was analyzed. The results showed that the flanking sequence was 299 bp,which was located on the third chromosome. The full length of the mutant gene was 5 746 bp,encoding 592 amino acid. The theoretical molecular weight of the encoding protein was 67. 5k Da,the hydrophobic amino acid content was 42. 06%,which had 10 transmembrane. It belonged to hydrophilic membrane protein. This result laid a foundation for the better use of Mutator to create new germplasm of sweet corn and it was very important for the development of endosperm and starch synthesis in maize.