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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (27): 37-44.

所属专题： 生物技术； 油料作物

• 农学 农业基础科学 • 上一篇    下一篇

花生Rab2基因的克隆及表达分析

李瑞 隋炯明   

  • 收稿日期:2012-03-05 修回日期:2012-04-22 出版日期:2012-09-25 发布日期:2012-09-25
  • 基金资助:

    山东省自然基金青年基金

Cloning and Expression Analysis of Rab2 Gene in Peanut

  • Received:2012-03-05 Revised:2012-04-22 Online:2012-09-25 Published:2012-09-25

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摘要：

Rab2是小GTP结合蛋白家族成员之一，控制着蛋白从内质网到高尔基体的运输，研究发现有些Rab2基因受多种逆境胁迫的诱导表达。为了获得花生Rab2基因，本试验根据拼接的cDNA序列设计1对特异性引物，通过RT-PCR克隆了1个花生Rab2基因。测序结果显示其cDNA含有长636 bp的完整开放阅读框，编码的蛋白含211个氨基酸，推测分子量为23.4 kDa。比对分析发现花生Rab2蛋白与多种生物的Rab2的氨基酸序列具有较高的同源性，荧光定量PCR研究表明AhRab2基因受低温、干旱、盐和植物激素ABA的诱导表达。这为利用基因工程手段调控花生的抗逆性奠定了基础。

关键词: 叶绿素荧光, 叶绿素荧光

Abstract:

Rab2 is a member of small GTP-binding protein family controlling protein transport from endoplasmic reticulum to golgi body, some of them are found to be responsive to different environmental stresses. In order to obtain Rab2 in peanut, according to the jointed EST sequences, a pair of specific primer was designed, and Rab2 gene from peanut (AhRab2) was amplified successfully by RT-PCR. After T-A cloning, sequencing result indicated that the cloned cDNA contained a continuous complete open reading frame (ORF) of 636 bp encoding a polypeptide of 211 amino acids with a calculated molecular mass of 23.4 kDa. Sequence comparison of the AhRab2 revealed high level amino acid sequence identity with Rab2 from other species. Transcription of AhRab2 gene was differentially regulated by different environmental stimuli such as cold, dehydration, NaCl and the plant hormone ABA using real-time PCR. This gene can be used to regulate stresses resistance of peanut by genetic engineering.