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龙眼成花逆转花芽α-tubulin基因的克隆与原核表达
Cloning and Prokaryotic Expression of α-tubulinGene in Longan Floral Reversion Buds
【摘要】 运用蛋白质组学方法比较龙眼(Dimocarpus longanLour.)正常成花和成花逆转花芽的差异蛋白质组,并应用RACE方法克隆其中上调表达的α-微管蛋白基因α-tubulin,获得一段长度为1641 bp的cDNA,其中包括1个1350 bp的开放阅读框[GenBank登录号:FJ479617(GI:218202929)]。将α-tubulin全长cDNA在大肠杆菌中表达,获得1个约49.6 kD的外源蛋白,经Western blotting验证为α-微管蛋白。RT-PCR和Western blotting分别检测了α-tubulin在转录和翻译水平上的表达,结果表明,α-微管蛋白在成花逆转的龙眼花芽中上调表达,可能是逆转花芽形态差异表现的原因之一。
【Abstract】 The differential proteins of floral reversion buds at different stages in longan(Dimocarpus longan Lour.) were compared using proteomics method.The results showed that α-tubulin could up-regulate in floral reversion buds of longan,and its gene α-tubulin was cloned using RACE method.A full length of 1641 bp cDNA,with a 1350 bp open reading frame,was obtained(GenBank accession number: FJ479617).When α-tubulin gene was transformed into E.coli and expressed,a 49.6 kD heterologous protein verified as α-tubulin by Western blotting was obtained.Different expression of α-tubulin at transcription and translation levels using RT-PCR and Western blotting,respectively,showed that α-tubulin up-regulated in longan floral reversion buds.It was one of reasons that floral reversion buds were different from normal flowering buds.
【Key words】 Longan; Flower bud; α-tubulin gene; Clone,Prokaryotic expression;
- 【文献出处】 热带亚热带植物学报 ,Journal of Tropical and Subtropical Botany , 编辑部邮箱 ,2011年01期
- 【分类号】S667.2
- 【被引频次】10
- 【下载频次】249