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作物学报 ›› 2009, Vol. 35 ›› Issue (2): 295-300.doi: 10.3724/SP.J.1006.2009.00295

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

青稞hblt4.2基因的克隆及功能分析

何涛1,2;贾敬芬2 *   

  1. 1青海大学生物科学系，青海西宁810016；2西北大学生命科学学院，陕西西安710069
  • 收稿日期:2008-05-26 修回日期:2008-10-24 出版日期:2009-02-12 网络出版日期:2008-12-11
  • 通讯作者: 贾敬芬
  • 基金资助:

    本研究由国家自然科学基金项目（30671082），陕西省教育厅重点项目（05JS48）资助。

Cloning and Function Analysis of hbltl4.2 Gene in Highland Barley(Hordeum vulgare L. var. nudum Hook. f.)

HE Tao1,2,JIA Jing-Fen2,*   

  1. 1Department of Biosciences, Qinghai University, Xining 810016,China; 2College of Life Sciences, Northwest University, Xi'an 710069,China
  • Received:2008-05-26 Revised:2008-10-24 Published:2009-02-12 Published online:2008-12-11
  • Contact: JIA Jing-Fen

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2083

被引次数

3

摘要：

根­据大麦blt14.2基因序列设计引物, 用青稞的DNA为模板扩增低温反应基因的全长序列, 命名为hblt14.2, 在GenBank登录号为EF514912。该基因含470 bp, 包括249 bp的开放阅读框、69 bp的5'非翻译区和152 bp的3'非翻译区, 编码82个氨基酸残基, 富含Gly、Ala、Leu和Val氨基酸, 与其他冷诱导基因编码蛋白具相似性。与大麦blt14.2基因具98.9%同源性, 所编码的蛋白存在2个氨基酸的差别。将hblt14.2基因连接CaMV35S启动子和NOS终止子后定向插入质粒pCAMBIA1301的多克隆位点中, 构建植物表达载体pCAMBIA-BI-hblt, 通过农杆菌介导转化烟草, 并对转基因烟草进行抗寒性检测。结果表明, 青稞hblt14.2基因与植物的抗寒性有一定关系。

关键词: 青稞, 低温, hblt4.2基因, 功能分析

Abstract:

The response to low temperature stress in highland barley indicates that there is a potential gene bank for cold resistance, which is helpful to develop the genes associated with resistance to cold and to survey the regulation mechanism. The objective of this study was to exploit a new gene with cold resistance, to validate its function. A pair of primer was designed based on the blt14.2 gene sequence documented in GenBank, a low-temperature-responsive (LTR) gene hblt14.2 was cloned by PCR from highland barley with GenBank accession number EF514912. Its length was 470 bp, containing 249 bp ORF, 69 bp 5'untranslated region (UTR) and 152 bp 3'UTR. Nucleotide sequence of hblt14.2 gene shares 98.9% (only five base pairs difference)homology with that of blt14.2 in barley, which encoded a protein of 82 amino acids. Theprotein was a highly hydrophilic small protein with rich Gly, Ala, Leu and Val amino acids, which were similar to proteins encoded by other LTR genes. The hblt14.2 ligated with CaMV 35S promoter and NOS terminator was inserted into multiple clone site in plasmid CAMBIA1301 to construct plant expression vector pCAMBIA-BI-hblt, which was transformed into tobacco mediated by Agrobacterium tumefaciens. Transgenic plants response to low temperature were investigated. The results suggested that the hblt14.2 is related to cold resistance in highland barley.

Key words: Highland barley, Low temperature, hbltl4.2 gene, Function analysis

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