CAO Peipei,MAO Yachao,LIU Tao,et al.Cloning and expression analysis of Cm14-3-3v gene in chrysanthemum[J].Journal of Nanjing Agricultural University,2017,40(5):820-826.[doi:10.7685/jnau.201612048]





Cloning and expression analysis of Cm14-3-3v gene in chrysanthemum
曹沛沛 毛雅超 刘涛 陈发棣 房伟民 陈素梅 蒋甲福
南京农业大学园艺学院, 江苏 南京 210095
CAO Peipei MAO Yachao LIU Tao CHEN Fadi FANG Weimin CHEN Sumei JIANG Jiafu
College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
chrysanthemumCm14-3-3v geneprotein structurestressgene expression
[目的]本文旨在克隆菊花‘神马’Cm14-3-3v基因,并初步分析其表达特征,为其功能研究奠定基础。[方法]从菊花‘优香’转录组库中获得编码拟南芥同源基因14-3-3v序列信息,根据NCBI比对确定基因的cDNA全长并进行克隆,再利用生物信息学方法对该蛋白的功能和活性位点进行预测,同时利用实时定量PCR分析了在非生物胁迫条件下该基因的表达变化。[结果]Cm14-3-3v的ORF序列长774 bp,编码257个氨基酸;亚细胞定位表明Cm14-3-3v定位在细胞质和细胞核;生物信息学分析表明该蛋白有19个Ser、7个Thr和7个Tyr潜在的磷酸化位点,在N-端有5个蛋白结合区;表达模式分析显示,Cm14-3-3v基因在根、茎、叶、花中均有表达,且受脱落酸(ABA)、茉莉酸甲酯(MeJA)、盐和冷胁迫的诱导表达。[结论]本文克隆得到菊花Cm14-3-3v基因,表达模式分析表明其受ABA、MeJA、盐和冷胁迫诱导表达。
[Objectives]We cloned chrysanthemum ‘Jimba’ Cm14-3-3v gene and analyzed its expression characteristics,which provided the foundation for functional research.[Methods]The sequence encoding the 14-3-3v was obtained based on the annotation of genes from a RNA seq library of chrysanthemum,and the full length cDNA was cloned with RT-PCR. The bioinformatics method was used to predict the function and the characteristics of the protein,and the expression levels of the gene under different abiotic stresses were analyzed by qRT-PCR.[Results]The ORF of Cm14-3-3v was 774 bp,encoding 257 amino acids. Subcellular localization analysis showed that Cm14-3-3v was located in the cytoplasm and nucleus. The protein had 19 Ser,7 Thr and 7 Tyr potential phosphorylation sites,5 protein-binding regions at the N-terminal. The expression pattern analysis showed that the Cm14-3-3v gene was expressed in roots,stems,leaves,flowers,and was induced by abscissic acid(ABA),methyl jasmonate(MeJA),salt and cold stress.[Conclusions]The expression of Cm14-3-3v gene in chrysanthemum was cloned,its expression was induced by ABA,MeJA,salt,cold stress.


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更新日期/Last Update: 2017-09-23