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农学学报

农学学报 ›› 2013, Vol. 3 ›› Issue (3): 4-9.

所属专题： 生物技术； 马铃薯

• 农艺科学 作物遗传育种 生理生化 • 上一篇    下一篇

甘薯Ran基因的克隆和表达分析

范乾程 王亚 隋炯明 毕英娜 刘媛 王晶珊   

  • 收稿日期:2013-02-01 修回日期:2013-02-26 出版日期:2013-03-20 发布日期:2013-03-20

Cloning and Expression Analysis of Ran Gene in Sweet Potato

  • Received:2013-02-01 Revised:2013-02-26 Online:2013-03-20 Published:2013-03-20

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摘要： Ran蛋白是小GTP结合蛋白家族成员之一，控制着蛋白和RNA分子的运输过程。研究发现有些Ran基因受多种逆境胁迫的诱导表达。为了获得甘薯Ran基因，本试验通过RT-PCR克隆了1个甘薯Ran基因。测序结果显示其含有长666 bp的完整开放阅读框，编码的蛋白含221个氨基酸，推测分子量为25.15 kDa。比对分析发现甘薯与多种生物的Ran蛋白的氨基酸序列同源性都超过90%，荧光定量PCR研究表明IbRan基因受低温、PEG、盐和植物激素ABA的诱导表达。这为利用基因工程手段调控甘薯的抗逆性奠定了基础。

关键词: 猪, 猪, 卵母细胞, 白藜芦醇, 抗冻能力, 基因表达

Abstract: Ran protein is a member of small GTP-binding protein family controlling the transport of protein and RNA, some of them are found to be responsive to different environmental stresses. In order to obtain Ran gene in sweet potato, according to the spliced EST sequences, a pair of specific primer was designed, and Ran gene from peanut (IbRan) 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 666 bp encoding a polypeptide of 221 amino acids with a calculated molecular mass of 25.15 kDa. Sequence comparison of the IbRan showed very high homology to the Ran subfamily from other species, with over 90% homology.Transcription of IbRan gene was differentially regulated by different environmental stimuli such as cold, PEG, NaCl and the plant hormone ABA using real-time PCR. This gene can be used to regulate stresses resistance of sweet potato by genetic engineering.