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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

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【作者】 游向荣黄榕辉王喜军黄春梅梁文裕陈伟

【Author】 YOU Xiang-rong 1a,1b,2,HUANG Rong-hui1b,WANG Xi-jun1b,HUANG Chun-mei1b,LIANG Wen-yu1b,CHEN Wei 1a,1b(1a.Key Laboratory of Ministry of Education for Genetics,Breeding and Multiple Utilization of Crops;1b.College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China;2.Institute of Agro-food Science and Technology,Guangxi Academy of Agricultural Sciences,Nanning 530007,China)

【机构】 福建农林大学作物遗传育种与综合利用教育部重点实验室福建农林大学生命科学学院广西农业科学院农产品加工研究所

【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.

【基金】 国家自然科学基金项目(30571293);福建省自然科学基金项目(2007J0045);教育部博士点基金(200803890009)资助
  • 【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
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