巴西橡胶树（Hevea brasiliensis）微管相关蛋白全长cDNA克隆及表达分析

Cloning and Identification of a cDNA Encoding Putative Microtubule-associated Proteins from Hevea brasiliensis

摘要:

从巴西橡胶树Hevea brasiliensis差减cDNA文库中分离到微管相关蛋白（Microtubule-associated protein,MAPs）基因片段,根据该基因片段序列信息,设计特异引物,采用cDNA末端快速扩增技术RACE（Rapid Amplification ofcDNA Ends）进行差异片段的5＇和3＇端的扩增,获得了长度为788bp的全长cDNA,该基因在GenBank中的登录号为AY461412。序列分析表明该基因包含完整的开放阅读框,编码144个氨基酸,与微管相关蛋白基因家族具有很高的同源性,推测该基因是微管相关蛋白基因。半定量RT-PCR检测证实它在胶乳中的表达强于叶中,胁迫处理（伤害及乙烯处理）使其表达上调。

Abstract:

The cell skeleton of laticiferous cell might play a very important role in latex production and latex flow, and in wound healing and laticifer blocking of tapped Hevea brasiliensis. In order to understand the relationship between cell skeleton of laticiferous cells and latex exploitation and laticifer blocking, a clone R329 was isolated from the subtracted cDNA library of Hevea brasiliensis, which showed high homology with microtubuleassociated proteins. The complete cDNA ofR329 was obtained by using 3＇-and 5＇-RACE strategies. It was 788 bp long containing an 435 bp OR_F, flanked by a 173 bp 5 ＇-UTR and a 179 bp 5 ＇-UTR including a poly（A） tail of 28 bp. The DNA sequence of R329 is available from the GenBank databases under the accession number AY461412. The transcripts of R329 were observed by RT-PCR, no signal was detected in leaves of Hevea brasiliensis, whereas significant signals of transcripts were clearly shown in latex isolated from virgin rubber trees, regularly tapped trees and ethylene-treated rubber trees. These results indicate that the expression of the gene is strongly induced by stress （wound and ethylene ） treatments.