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J Integr Plant Biol, 2001, 43 (9): 955-959, Research Article
Molecular Cloning of a Specific Expression Gene for Cold Acclimation in Cucumber
KANG Guo-Bin, XU Yong, YONG Wei-Dong, GE Lei, WANG Li-Ping, ZHANG Hai-Ying, WANG Yong-Jian and CHONG Kang
doi:
Abstract

A strategy of mRNA differential display was used for screening specific expression genes for cold acclimation in cucumber (Cucumis sativus L.). “Jinyan No.4”, a cold sensitive cultivar of cucumber, was treated under the conditions of 8 ℃ for 0, 6, 12, 24, 48 and 72 h, respectively, once it grew until two true leaves stage. A cold specific cDNA clone named as ccr18 was obtained. Data of differential display and Northern blot analysis showed that it expressed at cold acclimation for 12 h or more, and the longer duration at cold condition the higher it expressed. Southern blot assay suggested that it was a gene with single copy in chromosome of cucumber. Sequencing data showed the size of the cDNA clone was 639 bp in length. It shared 88% sequence with a BAC F14P3 genomic sequence of Arabidopsis thaliana chromosome Ⅲ at protein sequence level.

低温诱导的黄瓜ccr18 基因的cDNA 克隆及其表达特性分析
康国斌1 许勇1 雍伟东2 葛磊2王丽萍2 张海英1 王永健1 种康2*

(1. 国家蔬菜工程技术研究中心,北京100089;2. 中国科学院植物研究所分子与发育生物学研究中心,北京100093)

摘要:采用mRNA差异显示银染技术克隆得到在黄瓜(Cucumis sativus L.)冷敏型品种“津研4号”低温锻炼中特异表达基因的cDNA克隆(ccr18),其大小为639bp。在基因组中以单拷贝或低拷贝形式存在。Northern blot分析显示ccr18基因在12、24、48和72h低温处理的黄瓜中表达,在6h低温处理及对照中没有表达。这表明ccr18基因与黄瓜低温锻炼相关,序列同源性比较表明,它与拟南芥(Arabidopsis thaliana)染色体ⅢBAC库中的F14P3基因组序列具有88%的同源性。

关键词: 黄瓜;低温驯化; cDNA克隆;mRNA差异显示

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