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草地早熟禾PpGA20ox1基因的克隆及表达分析
Isolation and Expression Analysis of PpGA20ox1 Gene from Poa pratensis L.
【摘要】 利用RACE方法,从草地早熟禾叶片中克隆出1个GA20ox基因家族成员的cDNA序列,命名为PpGA20ox1,提交到Genbank,登录号为KX443595。PpGA20ox1开放阅读框长1095bp,编码含364个氨基酸的蛋白质序列;不含信号肽序列,为疏水蛋白,与黑麦草、小麦等物种的亲缘关系较近;用Gateway方法构建瞬时表达载体并注射烟草叶片观察亚细胞定位,发现其定位在细胞质和细胞核。用相对荧光定量qPCR方法研究基因的组织特异性及喷施赤霉素和烯效唑对基因表达的影响,结果表明PpGA20ox1基因在草地早熟禾叶片中表达量最高,根次之,叶鞘最少,且表达量受赤霉素和烯效唑抑制。
【Abstract】 A cDNA sequence of GA20 ox gene family,named as PpGA20ox1(Genebank accession number:KX443595),was isolated from the leaves of Kentucky bluegrass by RACE.The length of open reading frame was 1095 bp,encoding 364 amino acids.It’s hydrophobic protein,excluding signal peptide and was closely related with ryegrass and wheat.The transient expression vector was built by the method of Gateway and injected into the tobacco leaf to observe the subcellular location.The images indicated that this gene was located in the cytoplasm and nucleus.The tissue specific expression and effect of GA and UNI were conducted by Real-time qPCR.It showed that PpGA20ox1 expresses the highest in the leaf,next in the root,and in leaf sheath the least.PpGA20ox1 expression can be restrained by GA and UNI.
【Key words】 Kentucky bluegrass; PpGA20ox1 gene; Bioinformatics; Subcellular location; Expression analysis;
- 【文献出处】 中国草地学报 ,Chinese Journal of Grassland , 编辑部邮箱 ,2017年04期
- 【分类号】Q943.2;S688.4
- 【被引频次】5
- 【下载频次】172