摘要
以RT-PCR结合RACE方法克隆洋葱成花素同源基因AcLFT3的全长cDNA序列,利用生物信息学方法对蛋白结构进行预测分析,采用实时定量PCR法分析洋葱AcLFT3基因在不同时期不同器官的表达模式,为探讨洋葱抽薹开花的分子机制奠定基础。克隆获得的洋葱成花素同源基因AcLFT3 cDNA全长序列共838 bp,其中开放读码框为540 bp,编码由179个氨基酸组成、等电点为8.81、分子质量为20.1 ku的一个PEBP家族蛋白。AcLFT3与大葱成花素蛋白的同源性最高,为84.4%,其次为鸢尾。实时荧光定量PCR分析表明,AcLFT3基因在洋葱不同生长发育阶段器官中均有表达,以春化后抽薹前的叶片中表达量最高。
The full-length cDNA of florigen homologous gene AcLFT3 was obtained by RT-PCR and Rapid Amplification of cDNA Ends. The protein structure of AcLFT3 was analyzed by bioinformatics methods. The expression pattern of AcLFT3 among organs with different developmental stages in Allium cepa were analyzed by QRT-PCR. All the results will establish a foundation for molecular mechanism of bolting and florescence in Allium cepa. In this study, the cloned cDNA sequence of AcLFT3 in Allium cepa was 838 bp with a 540 bp open reading frame （ORF）. The deduced AcLFT3 protein was a member of the PEBP family, which had 179 amino acids. The molecular weight of AcLFT3 protein was 20.1 ku and its isoelectric point was 8.81. As AcLFT3 had the closest relationship to Allium fistulosum （84.4%）, followed by Iris fulva. QRT-PCR result showed that AcLFT3 was expressed during all the organs with different developmental stages. More interestingly, AcLFT3 was highly expressed in leaves of Allium cepa from the stage of vernalization to bolting.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2014年第9期40-46,共7页
Journal of Northeast Agricultural University
基金
黑龙江省科技特派员项目(GC13B809)
公益性行业(农业)科研专项项目(200903018)
关键词
洋葱
AcLFT3
荧光定量
序列分析
表达模式
Allium cepa
AcLFT3
fluorescence quantitative
sequence analysis
expression pattem
