东方百合T-DNA插入突变株侧翼序列的分离及分析

Flanking Sequences Isolation and Analysis of T-DNA Insertion into Oriental hybrid lily

采用反向PCR技术对东方百合(Oriental hybrid lily)T-DNA插入突变株的侧翼序列进行扩增并进行序列分析。结果表明:6个突变表型的植株都有T-DNA插入。扩增到的10个序列长度大都在500～900 bp左右。回收其中的8条亮带,克隆测序后,经NCBI BLASTx比对发现,除了B2序列因为太短没有显著的同源性以外,有1个是未知功能的蛋白。B1是Lys家族转录调节因子;CM1是C类细胞色素生成蛋白;CM2是糖转运跨膜蛋白;D1是特有的解旋酶家族;F1是rRNA小亚基甲基转移酶;G1是Arac家族转录调节因子。表明这些基因可能与这些突变表型有关。

The flanking sequences of T-DNA insertion mutants of Oriental hybrid lily（starfighter）were isolated by inverse PCR.A few noticeable phenotypes such as red leaf,toothed leaf,purple spot leaf,aclinic plant and lethal plants were observed.The inverse PCR analysis confirmed that there were T-DNA insertions for the six noticeable phenotype plantlets and ten flanking sequences of 500 to 900 bp length were obtained.Eight bright bands were recycled and sequenced.The predicted results of NCBI BLASTx showed that one was hypothetical protein,one had no significant similarity,B1 was transcriptional regulatory protein of LysR family,CM1 was the C-type cytochrome biogenesis membrane protein,CM2 was the permease of ABC sugar transporter,D1 was the distinct helicase family with a unique c-terminal domain including a metal-binding cysteine cluster,F1 was the putative rRNA small subunit methyltransferase,G1 was the Arac family transcriptional regulator.It was indicated that these flanking sequences might be correlated with these abnormal phenotypes.