Isolating the Mutator Transposable Element Insertional Mutant Gene mio16 of Maize Using Double Selected Amplification of Insertion Flanking Fragments (DSAIFF)
文献类型： 外文期刊
作者： Zhong Wen-juan 1 ; Zhang Mei-dong 1 ; Yang Liu-qi 2 ; Wang Ming-chun 2 ; Zheng Yong-lian 1 ; Yang Wen-peng 2 ; Gao You 1 ;
作者机构： 1.Huazhong Univ Agr, Natl Key Lab Crop Genet Improvement, Wuhan 430070, Peoples R China
2.Guizhou Acad Agr Sci, Inst Upland Food Crops, Guizhou Ctr Maize Engn Tech, Guizhou 550006, Peoples R China
关键词： maize (Zea mays L.);Mutator (Mu) transposable element;Mu flanking fragments (MFFs);double selected amplification of insertion flanking fragments (DSAIFF);mio16
期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：2.848； 五年影响因子：2.979 ）
ISSN： 2095-3119
年卷期： 2012 年 11 卷 10 期
页码：
收录情况： SCI
摘要： Mutator transposable element (Mu) has been used as an effective tool to clone maize (Zea mays L.) genes. One opaque endosperm mutant (mio16) was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments (DSAIFF), was employed to isolate the Mu flanking fragments (MFFs) of mio16. The target site duplications (TSDs) isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and mio16 was mapped in silica onto the physical position ranged from 229 965 021 to 229 965 409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of mio16 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of mio16 is alternated by Mu insertion.
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