龙眼成花逆转花芽α-tublin基因的克隆与原核表达

Cloning and Prokaryotic Expression of α-tublin Gene in Longan Floral Reversion Buds

摘要:

运用蛋白质组学方法比较龙眼(Dimocarpus longan Lour.)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.