为板栗FLC同源基因控制开花的相关功能及春化作用的相关分子机理提供研究基础,以罗田板栗为研究材料,分离并鉴定板栗花芽调控相关基因FLC,依据NCBI数据库提供的EST序列片段,利用RACE技术,从板栗叶片总RNA中分离了与开花相关的基因-CmFLCcDNA的序列,通过在线分析预测其序列同源性及3D结构,以pBluescript SK plus作为过渡载体构建其干扰表达载体。结果表明,该序列全长为968bp,其中有1个编码含227个氨基酸蛋白序列的开放阅读框。CmFLC蛋白的理论等电点为5.80,分子质量为25.62kDa,N端含有M盒保守序列,该蛋白的二级结构大部分由无规则卷曲和α-螺旋构成。CmFLC存在M盒、K盒两个特征性序列区域。CmFLC蛋白与毛白杨FLC蛋白的三维结构存在高度相似性。板栗的FLC蛋白属于植物进化分支,且与山核桃的FLC蛋白同属一支。设计特异引物扩增得到FLC基因的正、反义基因干扰片段,酶切结果显示插入片段大小为320bp,板栗FLC的干扰载体pc1301-ubi-CMFLCRNAi构建成功。结论:板栗中存在FLC同源基因,并可能参与板栗开花相关功能及春化调控。
From the NCBI database, 81,518 Lactuca sativa EST sequences were downloaded, from which 61,757 non-redundant sequences were obtained. In total, 2040 SSR loci were identified, with the frequency of 3.3%. Trinucleotide ...
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From the NCBI database, 81,518 Lactuca sativa EST sequences were downloaded, from which 61,757 non-redundant sequences were obtained. In total, 2040 SSR loci were identified, with the frequency of 3.3%. Trinucleotide repeat was dominant, followed by dinucieotide and hexanucleotide repeats. Among 181 types of repeat motifs, （AGA）n and （ATG）n were the dominant repeat motif types, taking up 12.01% and 3.53%, respectively. Totally, four material-special EST-SSR markers were characterized. This study enriches molecular markers of L. sativa, and also lays a theoretical foundation for following germplasm identification, molecular marker assisted breeding and genetic map construction.
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