华北农学报 ›› 2007, Vol. 22 ›› Issue (4): 9-13. doi: 10.7668/hbnxb.2007.04.003

所属专题: 杂粮作物 抗旱节水 生物技术

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谷子DnaJ蛋白基因的克隆

崔润丽1,2, 智慧1, 王永芳1, 李伟1, 李海权1, 黄占景3, 刁现民1   

  1. 1. 国家谷子改良中心, 河北省农林科学院谷子研究所, 河北 石家庄 050031;
    2. 河北化工医药职业技术学院, 河北 石家庄 050016;
    3. 河北师范大学生命科学学院, 河北 石家庄 050012
  • 收稿日期:2007-06-25 出版日期:2007-08-28
  • 通讯作者: 刁现民(1963-),男,河北南和人,博士,研究员,博士生导师,主要从事作物遗传育种与分子生物学研究。
  • 作者简介:崔润丽(1973-),女,河北鹿泉人,硕士,主要从事植物、微生物遗传和生物技术教学与研究工作。
  • 基金资助:
    国家自然科学基金(30630045);河北省自然科学基金资助(C2004000697)

Cloning of DnaJ-like Protein Gene from Foxtail Millet

CUI Run-li1,2, ZHI Hui1, WANG Yong-fang1, LI Wei1, LI Hai-quan1, HUANG Zhan-jing3, DIAO Xian-min1   

  1. 1. National Millet Improvement Center, Institute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050031, China;
    2. Hebei Chemical and Pharmaceutical College, Shijiazhuang 050016, China;
    3. College of Life Sciences, Hebei Normal University, Shijiazhuang 050016, China
  • Received:2007-06-25 Published:2007-08-28

摘要: DnaJ蛋白是近年研究较多的同热激等胁迫反应相关的基因。本研究用0.8%NaCl胁迫下的谷子幼苗提取RNA,用反转录酶M-MLV扩增得到第一链cDNA,再以第一链cDNA为模板进行PCR扩增,克隆获得了完整的谷子DnaJ蛋白基因,该基因cDNA全长1260 bp,编码419个氨基酸,具有DnaJ蛋白分子伴侣系统的3个保守结构域。将谷子DnaJ蛋白基因构建入表达载体pGFP,获得了谷子DnaJ基因的表达载体。该基因的克隆和表达载体构建,为谷子DnaJ蛋白基因的功能分析以及谷子耐热抗旱机理研究有重要意义。

关键词: 谷子, DnaJ, 基因克隆

Abstract: The DnaJ-like proteins and related genes are frequentily studied which are related with heatshock and other stresses in plant recentily. Using RT-PCR we cloned a foxtail millet Dna-J like gene in this report. RNA isolation was carried out by the Trizol method with 0.8%NaCl solution treated foxtail millet seedling; first chain cDNA was obtained with the reverse transcripte enzyme M-MLV and then used as templated to PCR cloning the cDNA sequence of the foxtail millet Dna-J like genes. A PCR fragment of predicted size of 1.2 kb was cloned and sequenced. The cloned fragment contains 1260 bp sequence which encodes a protein of 419 amino acids. Both the DNA sequence and amino acid sequence show great similarity with their corresponding sequences from rice, maize and wheat. The predicted protein has the three conserved domains which characteristic the structure of Dna-J protein family members. The cloned foxtail millet Dna-J gene was constructed into an expression vector for furthwer functional and transformation study.

Key words: Foxtail millet, Dna-J, Gene cloning

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引用本文

崔润丽, 智慧, 王永芳, 李伟, 李海权, 黄占景, 刁现民. 谷子DnaJ蛋白基因的克隆[J]. 华北农学报, 2007, 22(4): 9-13. doi: 10.7668/hbnxb.2007.04.003.

CUI Run-li, ZHI Hui, WANG Yong-fang, LI Wei, LI Hai-quan, HUANG Zhan-jing, DIAO Xian-min. Cloning of DnaJ-like Protein Gene from Foxtail Millet[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(4): 9-13. doi: 10.7668/hbnxb.2007.04.003.

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