Cloning,subcellular localization and spatiotemporal expression of VvGA2ox1 gene from grapevine | 葡萄VvGA2ox1基因克隆、亚细胞定位及时空表达分析
2013
Wang Xicheng, Nanjing Agricultural University,Nanjing(China),College of Horticulture | Wang Chen, Nanjing Agricultural University,Nanjing(China),College of Horticulture | Fang Jinggui, Nanjing Agricultural University,Nanjing(China),College of Horticulture
Chinese. 以‘藤稔’葡萄（Vitis vinifera×V.labrusca‘Fujiminori’）的花序、叶片和果实为试材,利用RT-PCR结合RACE技术,成功克隆了葡萄VvGA2ox1基因的cDNA序列,全长1 331 bp,共编码332个氨基酸。该基因在GenBank数据库的登录号为JQ608472。序列比对结果表明：VvGA2ox1与矮牵牛和棉花同源基因的氨基酸序列相似度分别为71.26%和71.08%。进一步构建了VvGA2ox1的亚细胞定位表达载体,定位结果显示,VvGA2ox1蛋白定位于细胞膜上。半定量RT-PCR与定量RT-PCR结果均表明,VvGA2ox1在葡萄花序、叶片和果实等器官中均有表达,但在花序、幼叶和小果中的表达水平较高。
Show more [+] Less [-]English. In this research,a full length cDNA of VvGA2ox1 was cloned from flowers,leaves and fruits of Vitis vinifera×V.labrusca ＇Fujiminori＇ using RT-PCR and rapid amplification of cDNA ends（RACE）.The full-length of the VvGA2ox1 gene was 1 331 bp,and an opening reading frame（ORF）encoding 332 amino acids.The sequence has been deposited in GenBank database with the accession number of JQ608472.Amino acid sequence analysis indicated that VvGA2ox1 shared 71.26% and 71.08% homology with Petunia hybrid Vilm and Gossypium hirsutum.The sub-cellular location analysis using expression vectors of VvGA2ox1 constructed showed that it was located in membrane.The semi-quantitative and fluorescent quantitative RT-PCRs were used to detect expression levels of VvGA2ox1 in different tissues of grapevine.The result showed that the VvGA2ox1 was expressed ubiquitously in vegetative and reproductive organs,such as flower,leaf and fruit.However,the expression level in flower,young leaf and small fruit was much higher.
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