油菜BnMKK4全长基因的克隆及表达分析

张腾国1,*, 王宁1, 王娟1, 王圆圆1, 张艳1, 孙万仓2, 常燕1, 夏惠娟3
1西北师范大学生命科学学院, 兰州730070; 2甘肃农业大学农学院, 兰州730070; 3兰州市第十二中学, 兰州730000

通信作者:张腾国;E-mail: zhangtg@nwnu.edu.cn;Tel: 0931-7971722

摘 要:

从‘陇油6号’油菜中克隆得到了一个新的BnMKK4基因的cDNA, 全长1 317 bp, 其中包括993 bp的开放阅读框, 159 bp 的5'非翻译区(5' UTR), 165 bp的3'非翻译区(3' UTR)。与拟南芥AtMKK4有很高的同源性, 因此命名为BnMKK4 (GenBank登 录号为JF268686)。该基因编码330个氨基酸的蛋白质, 分子量36.5 kDa, 等电点为9.01。实时荧光定量PCR结果显示该基因 表达受低温胁迫和盐胁迫的诱导, 表明该基因在油菜适应低温胁迫和盐胁迫的过程中发挥作用。

关键词:油菜‘陇油6号’; MKK4基因; 分子克隆; 表达分析

收稿:2012-01-04   修定:2012-04-16

资助:国家自然科学基金(30960065、31160089和31070358)和甘 肃省高等学校基本科研业务费项目。

Cloning and Expression Analysis of a BnMKK4 Gene from Brassica napus L.

ZHANG Teng-Guo1,*, WANG Ning1, WANG Juan1, WANG Yuan-Yuan1, ZHANG Yan1, SUN Wan-Cang2, CHANG Yan1, XIA Hui-Juan3
1School of Life Sciences, Northwest Normal University, Lanzhou 730070, China; 2College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China; 3Lanzhou No. 12 Middle School, Lanzhou 730000, China

Corresponding author: ZHANG Teng-Guo; E-mail: zhangtg@nwnu.edu.cn; Tel: 0931-7971722

Abstract:

In this study, we isolated a novel MAPKK gene, BnMKK4, from Brassica napus. The cloned fulllength cDNA was 1 317 bp, contained a 993-bp opening reading frame (ORF), a 5'-untranslated region (5' UTR) of 159 bp, and a 3'-untranslated region (3' UTR) of 165 bp. The BnMKK4 exhibited closest homology to AtMKK4, so it was named the BnMKK4 (GenBank accession No. JF268686). The deduced protein was 330 amino acids with molecular weight 36.5 kDa and isoelectric point 9.01. RT-PCR analysis revealed that the transcript level of BnMKK4 was higher in experimental plants than in control plants. The results suggested that the BnMKK4 gene is involved in response to cold and salt stresses and that BnMKK4 played an important role during cold and salt stresses in Brassica napus.

Key words: Brassica napus L. ‘Longyou 6’; MKK4 gene; molecular cloning; expression analysis

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