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采用TAIL-PCR技术从经鉴定含Ac/Ds双元件的材料中扩增Ds侧翼序列并测序,对水稻Ac×Ds后代基因组DNA进行Ac和Ds插入的PCR分析。利用NCBI的BLAST软件,以Ds侧翼序列为待查询序列进行GenBank在线搜索比对,获得Ds插入相关基因的染色体定位和功能注释等信息。对扩增的93个有效Ds侧翼序列进行分析,结果显示,有21个水稻杂交后代中Ds插入于基因编码区,其余72个插入在基因间序列,其中12个插入在特定基因的上游3kb以内的间隔区。本研究强调了提高Ds侧翼序列扩增和Ac/Ds植株筛选效率的技术关键。
TA-PCR was used to amplify the Ds flanking sequence from the identified Ac / Ds-containing double-stranded material and sequenced. PCR analysis of Ac and Ds insertions in the genomic DNA of Ac × Ds progeny of rice was carried out. Using NCBI BLAST software, the Ds flanking sequences were searched for GenBank online searching and aligning to get the information about chromosome location and function annotation of Ds insertion related genes. Analysis of 93 valid Ds flanking sequences showed that Ds was inserted into the coding region of the gene in 21 hybrid rice progenies and the remaining 72 were inserted in the intergenic sequence, of which 12 were inserted within 3kb upstream of the specific gene Of the compartment. This study emphasizes the key technology to improve the efficiency of Ds flanking sequence amplification and Ac / Ds plant selection.