金银花叶绿素a/b结合蛋白基因LjCab克隆与表达特性分析

Cloning and expression characterization of LjCab gene of the light harvesting chlorophyll a/b binding protein fromLonicera japonica Thunb.

采用设计简并性引物和RACE克隆相结合,从金银花中克隆了捕光叶绿素a/b结合蛋白基因全长,命名为LjCab(GenBank号:JQ621945.1).基因全长为960bp,开放阅读框为798bp,其编码265个氨基酸,基因不含内含子属于光系统II的Cab基因.对LjCab基因核苷酸序列进行亲缘关系分析发现其与烟草和五彩椒的Cab基因亲缘关系较近.金银花在不同条件处理下,对LjCab基因进行定量PCR检测分析,结果显示:LjCab基因受光、6-BA、GA3和NAA诱导表达,但是黑暗和ABA激素处理对LjCab基因的表达没有明显的影响.本研究为进一步解析金银花光合作用机理及其相关基因的诱导表达特性提供了参考.

In this study,a gene,named as LjCab（GenBank accession number：JQ621945.1）was cloned and characterized using degenerate primer design and RACE cloning technology,which encoded light harvesting chlorophyll a/b binding protein in Lonicera japonica Thunb..The full length of LjCabis 960 bp,in which the open reading frame that encodes 265 amino acids is 798 bp.This gene belongs to photosystem Ⅱwithout intron.DNA sequence conservation analysis showed that the sequence of LjCab had high identity with previously cloned Cab gene of Nicotiana tabacum or Capsicum annuum.Quantitative PCR demonstrated that the expression of LjCab significantly increased under light,or with 6-BA,GA3 or NAA treatment,while no response at transcriptional stage under darkness or with ABA treatment.These results provided references for better understanding on the mechanism of photosynthesis and gene expression in Lonicera japonica Thunb.