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园艺学报 ›› 2020, Vol. 47 ›› Issue (3): 581-589.doi: 10.16420/j.issn.0513-353x.2019-0357

• 研究报告 • 上一篇    下一篇

铁皮石斛亲环蛋白基因 DoCyP 的克隆及表达分析

付亚娟 1,2,陈霞婷 1,乔 洁 1,2,王 晶 1,2,李文静 1,侯晓强 1,2,*   

  1. 1 廊坊师范学院生命科学学院,河北廊坊 065000;2 河北省食药用菌资源高值利用技术创新中心,河北廊坊 065000
  • 出版日期:2020-03-25 发布日期:2020-03-25
  • 基金资助:
    河北省教育厅青年拔尖人才项目(BJ2016045);国家自然科学基金青年基金项目(31100314);廊坊师范学院博士基金项目(LSLB201405);河北省高等学校遗传学重点发展学科项目(201221)

Molecular Cloning and Expression Characterization of Cyclophilin Gene(DoCyP)in Dendrobium officinale

FU Yajuan1,2,CHEN Xiating1,QIAO Jie1,2,WANG Jing1,2,LI Wenjing1,and HOU Xiaoqiang1,2,*   

  1. 1College of Life Science,Langfang Normal University,Langfang,Hebei 065000,China;2Technical Innovation Center for Utilization of Edible and Medicinal Fungi in Hebei Province,Langfang,Hebei 065000,China
  • Online:2020-03-25 Published:2020-03-25

摘要: 采用 RT-PCR 技术克隆了铁皮石斛(Dendrobium officinale Kimura et Migo)亲环蛋白基因(DoCyP),其大小为 522 bp,编码 173 个氨基酸。生物信息学预测,DoCyP 相对分子质量 18.25 kD,理论等电点 8.72,含肽脯氨酰顺反异构酶保守结构域,无跨膜结构域,是稳定的亲水性蛋白质。SOPMA 预 测 DoCyP 的二级结构主要由无规卷曲和延伸链构成。系统进化树揭示铁皮石斛 DoCyP 与深圳拟兰 CyP聚为一支,亲缘关系较近。qRT-PCR 分析表明,DoCyP 在铁皮石斛组织中为组成型表达,在根中表达量最高,叶和茎中表达量较低。4 ℃胁迫 12 h,DoCyP 在铁皮石斛根和茎中表现为上调表达,在叶中下调表达。推测 DoCyP 参与铁皮石斛对低温胁迫的应答。

关键词: 铁皮石斛, 亲环蛋白, DoCyP, qRT-PCR, 低温胁迫

Abstract: The cyclophilin gene(DoCyP)in Dendrobium officinale was cloned by RT-PCR. The full-length of DoCyP was 522 bp,encoding 173 amino acids. Bioinformatics predicted that DoCyP is a stable hydrophilic protein with a relative molecular mass of 18.25 kD,containing a peptide-prolyl cis-transisomerase domain. The theoretical isoelectric point of DoCyP is 8.72,and no transmembrane domain. SOPMA predicted that the secondary structure of DoCyP primarily consists of random coils and extended chains. The phylogenetic tree indicated that DoCyP of D. officinale has a relatively close genetic relationship with Apostasia shenzhenica CyP. qRT-PCR results revealed that DoCyP was constitutively expressed in different tissues of D. officinale. The relative expression of DoCyP was higher in roots,compared to leaves and stems. Under low-temperature(4 ℃)stress,the expression of DoCyP in different D. officinale tissues showed different trends. Compared with normal expression level,DoCyP expression after treatment with 4 ℃ for 12 h was up-regulated in roots,while down-regulated in stems and leaves,suggesting that DoCyP may take part in response to low temperature stress in D. officinale.

Key words: Dendrobium officinale, cyclophilin, DoCyP, qRT-PCR, low temperature stress

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