金银花丙酮酸激酶基因的克隆与表达特性分析
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贵州省科技计划厅联合基金项目(黔科合LH字[2014]7219);贵州省教育厅重点项目(黔教合KY字[2014]281)


Cloning and Expression Characterization of Gene Encoding Pyruvate Kinase from Lonicera japonica Thunb.
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    摘要:

    采用RT-PCR和RACE技术,首次从金银花中克隆丙酮酸激酶基因全长cDNA,命名为LjPkc(GenBank登录号JQ621946.1),LjPkc基因编码区序列全长为1 533 bp,共编码510个氨基酸。基于基因序列亲缘关系分析结果显示,LjPkc基因与烟草Pkc基因亲缘关系最近,属于双子叶植物Pkc基因家族。蛋白质亚细胞定位分析认为LjPkc在线粒体或叶绿体中发挥作用。LjPkc蛋白质的三维空间结构预测结果显示,LjPkc蛋白主要由α-螺旋、不规则盘绕组成。定量PCR检测发现,LjPkc基因在金银花不同组织中广泛表达,但表达水平各有差异,黄花中表达量最高。该研究丰富了Pkc基因库资源,为揭示LjPkc基因在金银花糖代谢方面的生物学作用奠定了基础。

    Abstract:

    A full-length cDNA encoding pyruvate kinase was cloned firstly by RT-PCR and RACE-PCR from Lonicera japonica Thunb.,designated LjPkc (GenBank accession number JQ621946.1).The length of coding sequence (CDS) is 1 533 bp,which codes for a protein with 510 amino acids.LjPkc displayed high sequence identity with previously cloned Pkc gene of Nicotiana sylvestris and was clustered close to Pkc family genes of dicotyledon plants.Subcellular localization analysis showed that LjPkc protein may play an important role in the chloroplasts or mitochondria.The LjPkc proteins mainly have alpha-helix and random coil by spatial structure analysis.qPCR analyses showed that LjPkc is widely expressed in stem,leaf and flower of L.japonica Thunb.,but the mRNA of LjPkc has the highest expression level in yellow flower.The research is not only to enrich the gene resource of Pkc gene family,but also lay a foundation to reveal the biology function of LjPkc in sugar metabolism.

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蒋向辉,冯仕彪.金银花丙酮酸激酶基因的克隆与表达特性分析[J].西北植物学报,2015,35(12):2373-2378

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  • 在线发布日期: 2016-01-11
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