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园艺学报 ›› 2009, Vol. 36 ›› Issue (4): 521-526.

• 蔬菜 • 上一篇    下一篇

马铃薯C-8,7甾醇异构酶基因(StSI1) cDNA克隆及表达

牛洪斌1;白润娥2;邓德芝1;尹 钧1*   

  1. (1河南农业大学国家小麦工程技术研究中心, 郑州450002; 2河南农业大学植物保护学院, 郑州450002)
  • 收稿日期:2009-01-19 修回日期:2009-03-31 出版日期:2009-04-25 发布日期:2009-04-25
  • 通讯作者: 尹 钧

Cloning and Expression of C-8,7 Sterol Isomera se Gene (StSI1) cDNA fromPotato

NIU Hong-bin1,BAI Run-e2,DENG De-zhi1,and YIN Jun1*   

  1. (1National Engineering Research Center forW heat, Henan AgriculturalUniversity, Zhengzhou 450002, China; 2College of PlantProtection, Henan Agricultural University, Zhengzhou 450002, China)
  • Received:2009-01-19 Revised:2009-03-31 Online:2009-04-25 Published:2009-04-25
  • Contact: YIN Jun

摘要: 以拟南芥C-8, 7甾醇异构酶的氨基酸序列为信息探针搜索GenBank数据库, 对高度同源的马
铃薯EST序列进行拼接、引物设计和RT2PCR扩增, 扩增产物测序结果证实获得一个马铃薯C-8, 7甾醇异构酶基因(StSI1) 的全长cDNA序列。序列分析结果显示, StSI1全长886 bp, 包含59 bp的5′非编码序列、161 bp的3′非编码序列和一个长度为666 bp编码221个氨基酸的开放阅读框, 分子量约为25 kD。氨基酸结构分析显示该蛋白的N端含有一个长度由35个氨基酸残基组成的信号肽, C端成熟肽区域含有典型的类EBP结合域。氨基酸比对分析表明, StSI1与已知C-8,7甾醇异构酶同源性介于32.9% ~61.3%之间, 与拟南芥AtSI1相似性最(61.3% ) 。RT-PCR 表达谱分析显示, StSI1在马铃薯的块茎芽眼和表皮组织中均能表达, 并且该基因的表达水平受贮藏温度升高和光照增强的正向调节。

关键词: 马铃薯, 基因克隆, 表达分析

Abstract: C-8,7 sterol isomerase plays a vital role in the sterol biosynthesis pathway. UsingA rabidopsis C-8, 7 sterol isomerase (GenBank accession No. AAD03489) amino acid sequence as a querying probe,many highly homologous EST sequenceswere obtained from GenBank and a putative cDNA sequence of potato C-8, 7 sterol isomerase was assembled. Futhermore, a putative C-8, 7 sterol isomerase gene cDNA, named StSI1 (GenBank accession No. EU103613 ) was cloned from potato (Solanum tuberosum L. ). StSI1 was 886 bp in full length, including a 5′untranslated region of 59 bp, 3′untranslated region of 161 bp with poly A, and a 666 bp length open reading frame (ORF) encoding 221 amino acids with the molecular weight of 25 kD. Software prediction results showed that StSI1 contained a 35 amino acid lenghth signal pep tide at the N terminal. Homology analysis showed that the deduced amino acid sequence of StSI1 shared 32.9% - 61.3% identity with sterol isomerase from other organisms. RT2PCR analysis showed that the StSI1 was highly expressed in skin and eyes of potato stem tuber. The exp ression profiling also showed that StSI1 mRNA was up regulated intensively both by temperature and light.

Key words: potato, gene cloning, expressing analysis

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