银中杨DREB基因的克隆及表达  

Cloning and expression of DREBgene from Populus albaxp

作　　者：翟俊峰[1] 王法微[2] 王南[2] 张闯[1] 迟晓娟[1] 宗俊梅[1] 李海燕[1,2] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118

出　　处：《西北农林科技大学学报（自然科学版）》2012年第6期79-85,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(30971804);国家转基因生物新品种培育重大专项"基因表达调控技术研究"(2010ZX08010-002);教育部新世纪优秀人才计划项目(NCET-08-0693);长春市科技局计划项目(09YJ24)

摘　　要：【目的】克隆银中杨抗逆转录因子DREB基因,为其功能的深入研究奠定基础。【方法】根据DREBAP2/EREBP保守区设计1对简并引物,采用PCR方法对银中杨(Populus albaxp)转录因子DREB基因中间片段进行克隆;再利用反向PCR方法对该基因的旁侧序列进行克隆,最终将两侧序列与中间片段拼接得到基因全长;同时,根据基因序列设计1对特异引物,利用荧光定量PCR分析银中杨DREB基因在不同逆境胁迫中的表达情况。【结果】成功地从银中杨中克隆到1个DREB基因,命名为PaDREB(注册号:EF582843)。该基因序列长935bp,ORF为819bp,编码272个氨基酸;同源性比较与系统发育分析证实,银中杨DREB属于DREB转录因子家族,并与月季DREB2C具有较高的同源性;荧光定量PCR检测结果显示,盐、低温、干旱及ABA均能诱导DREB基因的表达。【结论】首次从银中杨中分离并克隆了DREB基因,并证实其参与了银中杨对盐、低温、干旱及ABA的应答。【Objective】 The study was done to clone the stress induced DREB transcription factor gene from Populus albaxp and provide a starting point for the further studies in functional analysis.【Method】 We firstly cloned the partial fragment of DREB gene from Populus albaxp using homology cloning methods,according to AP2/EREBP conserved sequences.Then,the flanking sequence of DREB gene was obtained using reverse PCR.The flanking sequences and the partial fragment were combined to a full length sequence,named PaDREB.A real-time PCR was performed to observe the expression profiling of PaDREB under various stresses using a pair of specific primers.【Result】 We obtained the full length of PaDREB gene and submitted to Genbank（Accessed Number：EF582843）.The full-length of cDNA is 935 bp and the open reading frame（ORF） is 819 bp,encoding 272 amino acids;Phylogenetic trees showed that PaDREB belongs to the DREB super family and shared a higher identity with the DREB2C.According to real-time PCR analysis,we found that salt,low temperature,drought and ABA could all induce the expression of PaDREB dramatically.【Conclusion】 We first cloned PaDREB gene from Populus albaxp.The results of RT-PCR showed that PaDREB gene could be involved in the response of salt,low temperature,drought and ABA treatment.

关 键 词：银中杨 DREB转录因子 基因克隆 基因表达 

分 类 号：S792.119.04[农业科学—林木遗传育种]