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植物研究 ›› 2008, Vol. 28 ›› Issue (6): 705-709.doi: 10.7525/j.issn.1673-5102.2008.06.013

• 论文 • 上一篇    下一篇

京大戟hmgr基因家族3个保守区片段的克隆与分析

曹小迎1;蒋继宏1*;鞠秀云1;戴传超2   

  1. (1.徐州师范大学江苏省药用植物生物技术重点实验室,徐州 221116) (2.南京师范大学生命科学学院,南京 210097)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-11-20 发布日期:2008-11-20
  • 通讯作者: 蒋继宏
  • 基金资助:
     

Cloning and Analysis of hmgr Gene conserved fragments in Euphorbia pekinensis Rupr

CAO Xiao-Ying;JIANG Ji-Hong*;JU Xiu-Yun;DAI Chuan-Chao   

  1. (1.Xuzhou Normal University,Key Laboratory of Biotechnology for Medicinal Plant,Xuzhou 221116) (2.College of Life Sciences,Nanjing Normal University,Nanjing 210097)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-11-20 Published:2008-11-20
  • Contact: JIANG Ji-Hong
  • Supported by:
     

摘要: 采用RT-PCR技术以及两条简并引物,以京大戟嫩叶总RNA反转录的cDNA为模板扩增3-羟基-3-甲基戊二酰辅酶A还原酶基因的保守区片断。序列分析表明,所克隆的cDNA保守区序列长度为458 bp,而且同时得到了三个不同的核苷酸序列,分别命名为hmgr1、hmgr2、hmgr3。与之前得到的京大戟hmgr保守区片段的核苷酸序列的同源性分别为98.03%、96.29%、78.38%,推断的相应氨基酸序列的同源性分别为98.68%、96.71%和85.53%。推断这可能是该基因家族中的三个新的成员。而且同源序列比对发现,由这三个核苷酸序列推断的氨基酸序列与其它植物都有较高的同源性。种系进化树分析表明hmgr3与hmgr1、hmgr2及以前报道的京大戟的hmgr之间有较大的差别。

关键词: 京大戟, 3-羟基-3-甲基戊二酰辅酶A还原酶, 基因克隆

Abstract: Based on total RNA extracted from Euphorbia pekinensis, a 458 bp fragment of hmgr was obtained using reverse transcription PCR strategy and two degenerated oligonucleotides. Sequence analysis revealed that the conserved fragments are 458 bp. At the same time, the three fragments named hmgr1,hmgr2 and hmgr3 were obtained, which were 98.03%、96.29% and 78.38% identification in nucleotide acid compared with hmgr in E.pekinensis reported previously, and 98.68%, 96.71% and 85.53% identification in corresponding amino acid. The three fragments may be new members of hmgr gene family. Sequencing analysis also showed that the three framents had high identity with hmgr from other plants. Deduced amino acid sequence were also analyzed by PROSITE, ClustalX and Phylogenic tree, and data indicated that hmgr3 were different from hmgr1, hmgr2 and hmgr in E.pekinensis reported previously.

Key words: Euphorbia pekinensis Rupr, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR), gene cloning

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