菊花MADS-box基因的克隆与表达载体构建

doi:10.7668/hbnxb.2016.03.004

摘要/Abstract

摘要： 为了给菊花CmFUL基因功能鉴定与遗传改良奠定基础,采用RT-PCR法从菊花叶片中克隆出1个MADS-boxA类基因的编码区序列,命名为CmFUL(GenBank登录号KT894379)。CmFUL基因编码区ORF片段长度为741bp,编码246个氨基酸。CmFUL蛋白属于亲水性蛋白,预测该蛋白内含12个磷酸化位点和2个潜在的N-糖基化位点。氨基酸序列比对和系统进化树分析表明,CmFUL蛋白与CMD41蛋白的相似性为95.6%,同属于AP1/FUL亚家族的FUL-like进化枝。实时荧光定量PCR分析表明,CmFUL基因在花期不同组织中均有表达,但表达丰度不一。在花蕾中表达量最高,其次是管状花,根和舌状花中痕量表达;同一朵花中管状花的表达量约为舌状花中的12倍。分析认为,CmFUL可能在促进开花及子房建成中起重要作用。将CmFUL基因连接到pCAMBIA1300载体上,成功构建了高效植物表达载体。

关键词: 菊花, FUL-like, 基因克隆, 表达分析, 生物信息学

Abstract: In order to research the identification of gene function and genetic improvement in Chrysanthemum,a MADS-box family class A gene was isolated from the leaf of Chrysanthemum morifolium by using RT-PCR method,which was designated CmFUL gene(The accession number in GenBank:KT894379).The coding region of this gene(ORF) was 741 bp and encoded 246 putative amino acid residues.Encoding product of CmFUL gene was a kind of hydrophilic protein and predicted that the protein contained 12 phosphorylation sites and 2 potential N-Glycosylation sites.Sequence and phylogenic analyses revealed that CmFUL was close to CDM41 with 95.6% sequence similarity which both belonged to the FUL -like clade of AP1/FUL subfamily.The expression analyses of CmFUL by Real-time fluorescent quantitative PCR indicated that this gene was expressed in different parts in bloom stage,but the expression abundance was varied widely.The highest amount of expression was observed in buds,followed by in tubiform florets.The expression level in roots and lingulate florets were lowest.It was worth noting that the level expressed in tubiform florets was 12 times of that in lingulate florets between a same flower.The results implied that CmFUL might play an important role in promoting flowering and ovary development.CmFUL was linked to pCAMBIA1300 vector and the plant expression vector was constructed successfully.

Key words: Chrysanthemum morifolium, FUL -like, Gene cloning, Expression analysis, Bioinformatics

中图分类号:

Q949.71+8.43

梁芳, 许申平, 蒋素华, 袁秀云, 崔波, 张静. 菊花MADS-box基因的克隆与表达载体构建[J]. 华北农学报, 2016, 31(3): 25-31. doi: 10.7668/hbnxb.2016.03.004.

LIANG Fang, XU Shenping, JIANG Suhua, YUAN Xiuyun, CUI Bo, ZHANG Jing. Cloning,Expression and Vector Construction of a MADS-box Gene in Chrysanthemum morifolium[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(3): 25-31. doi: 10.7668/hbnxb.2016.03.004.

http://www.hbnxb.net/CN/Y2016/V31/I3/25

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