草莓多酚氧化酶基因全长的克隆及表达载体构建  

Cloning of strawberry polyphenol oxidase genes and construction of their expression vectors

作　　者：张煜晗 张希[1] 董莉[1] 董清华[1] ZHANG Yuhan;ZHANG Xi;DONG Li;DONG Qinghua(Plant Science and Technology College,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院植物科学技术学院/北京林果业生态环境功能提升协同创新中心,北京102206

出　　处：《北京农学院学报》2018年第2期7-14,共8页Journal of Beijing University of Agriculture

基　　金：科技创新服务能力建设-协同创新中心-林果业生态环境功能提升协同创新中心(市级)(PXM2017-014207-000043);北京市教委科技发展项目(KM200710020012);北京市自然科学基金重点项目(6171001)

摘　　要：【目的】为了提高草莓的果实品质和经济价值。【方法】利用改良CTAB法提取五叶草莓叶片总DNA,反向PCR(IPCR)方法克隆基因。【结果】得到了2条完整的草莓多酚氧化酶基因全长3 591bp和1 809bp,分别编码1 196和602个氨基酸,分别命名为FpPPO1与FpPPO_2,GenBank登录号分别为HM063946和HM063947。FpPPO1和FpPPO_2与其他物种中的多酚氧化酶基因比较,氨基酸相似性分别为57%~77%与74%~99%。最后成功构建了多酚氧化酶基因反义植物表达载体pCP430和正义植物表达载体pCP1809。【结论】克隆得到草莓FpPPO1和FpPPO_2基因全长并构建转基因载体,为研究草莓中多酚氧化酶基因的功能奠定了基础。【Objective】To further clarify roles of polyphenol oxidase(PPO)in strawberry is the objective of this study.【Methods】The total DNA was extracted from the leaves of Fraxinus mandshurica by a modified CTAB method,and reverse transcription PCR(RT\|PCR)was used to clone the gene.【Results】The full lengths of two strawberry PPO genes were 3 591 bp and 1 809 bp,encoding 1 196 and 602 amino acids,and named as FpPPO1(GenBank No.HM063946)and FpPPO2(GenBank No.HM063947),respectively.The amino acid sequences of FpPPO1 and of FpPPO2 shared 57%~77%and 74%~99%identity with that of other plants,respectively.Finally,the antisense plant expression vector pCP430 and the sense plant expression vector pCP1809 of the PPO genes were successfully constructed.【Conclusion】In this study,the full length of FpPPO1 and FpPPO2 genes were cloned and the transgenic expression vectors were constructed.These rusults lay a foundation for the study of function of the polyphenol oxidase genes.

关 键 词：草莓 多酚氧化酶 基因克隆 

分 类 号：S668.4[农业科学—果树学]