黄瓜 CsCCD7 基因的克隆及表达研究

Cloning of CsCCD7 and Its Expression in Cucumis sativus L.

摘要:

使用RACE技术克隆黄瓜(Cucumis sativus L.)腋芽生长抑制基因并进行生物信息学和半定量RT-PCR分析。结果表明:从黄瓜腋芽中成功克隆了拟南芥(Arabidopsis thaliana)AtCCD7/MAX3同源基因,命名为CsCCD7(GenBank登录号:HQ005419);CsCCD7基因序列含有1665 bp的开放阅读框(ORF),编码554个氨基酸;编码的蛋白质命名为CsCCD7,隶属于CCD蛋白家族成员,蛋白质的二级结构和三级结构预测显示其富含β折叠和β转角以及无规卷曲,是不稳定蛋白。CsCCD7在根中的表达量最高,在多分枝、矮化黄瓜D0462中的表达量最低,这说明CsCCD7蛋白可能参与调控植物分枝信号的转导及分枝相关基因的表达调控。

关键词:黄瓜;腋芽生长抑制基因;基因克隆;生物信息学分析;基因表达

Abstract:

The cucumber (Cucumis sativus L.) inhibiting axillary bud growth gene, CsCCD7, had been cloned (GenBank accession number: HQ005419) using by RACE, and its bioinformatics and semi-quantitative RT-PCR analysis were studied. The results showed that the CsCCD7 was the homologue gene of AtMAX3/CCD7 in Arabidopsis thaliana, which contain ORF with 1665 bp, and code the protein with 554 amino acids, named CsCCD7, belonging to CCD protein family. The secondary and tertiary structure prediction of CsCCD7 showed having rich β sheets, β turns and irregular curls, which indicated it was unstable protein. CsCCD7 expression in roots was the highest, while that in dwarf cucumber 'D0462’ with lot of branches were the lowest by RT-PCR analysis. It suggested that the CsCCD7 protein could revolve in control branch signal transduction and related gene expression in cucumber.

Key words:Cucumber;Inhibiting axillary growth gene;Gene cloning;Bioinformatics analysis;Gene expression