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植物研究 ›› 2008, Vol. 28 ›› Issue (4): 412-416.doi: 10.7525/j.issn.1673-5102.2008.04.007

• 论文 • 上一篇    下一篇

RT-PCR克隆甜菜硝酸还原酶cDNA全长序列及分析

张 杰1,2;彭胜民1;周 波2;战晴晴2;张荣沭2;马凤鸣1*   

  1. (1.东北农业大学农学院,哈尔滨 150030) (2.东北林业大学生命科学学院,哈尔滨 150040)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-07-20 发布日期:2008-07-20
  • 通讯作者: 马凤鸣
  • 基金资助:
     

Cloning and Sequence Analysis of Nitrate Reductase Gene from Beta vulgaris by RT-PCR

ZHANG Jie;PENG Sheng-Min;ZHOU Bo;ZHAN Qing-Qing;ZHANG Rong-Shu;MA Feng-Ming*   

  1. (1.College of Agronomy,Northeast Agricultural University,Harbin 150030) (2.College of Life Sciences,Northeast Forestry University,Harbin 150040)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-07-20 Published:2008-07-20
  • Contact: MA Feng-Ming
  • Supported by:
     

摘要: 根据GenBank中已公布的甜菜(Beta vulgaris)硝酸还原酶(nitrate reductase)基因序列(gb∣ABW05098.1∣),设计引物,以50 mmol·L-1 KNO3溶液处理的甜菜幼苗为材料,从总RNA中通过RT-PCR分离得到一个硝酸还原酶基因,其cDNA长2 760 bp,包含了完整的基因编码序列,与已公布的硝酸还原酶基因序列相似性达99%。Southern杂交分析表明,硝酸还原酶基因在甜菜基因组中可能以两个拷贝或低拷贝形式存在。根据其编码的氨基酸序列,利用生物信息学预测了其亚细胞定位和蛋白质的三级结构。

关键词: 甜菜, 硝酸还原酶, 基因克隆, RT-PCR

Abstract: A pair of primers was designed according to the sequence of nitrate reductase gene in GenBank (gb∣ABW05098.1∣). The homolog cDNA with 2 760 base pairs of nitrate reductase gene was obtained by RT-PCR from total RNA using Beta vulgaris treated by 50 mmol·L-1 KNO3 as material. DNA sequence analysis showed that it contains the entire open reading frames that encode deduced protein with 905 amino acid residues and shows 99% sequence identity to B. vulgaris nitrate reductase gene. Southern hybridization experiments revealed that nitrate reductase gene might be exist as two copy or low copy in the genome of B. vulgaris. According to the amino acid sequences, the subcellular location and structure of the nitrate reductase were predicted.

Key words: Beta vulgaris, nitrate reductase, Gene cloning, RT-PCR

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