大豆CHI1基因启动子的克隆及功能初探  

Molecular Cloning and Function Analysis of a GmCHI1 Promoter

作　　者：赵春梅[1] 范习 薛仁镐[1] 

机构地区：[1]青岛农业大学生命科学学院山东省高校植物生物技术重点实验室,山东青岛266109

出　　处：《华北农学报》2017年第4期32-36,共5页Acta Agriculturae Boreali-Sinica

基　　金：转基因生物新品种培育重大专项资助项目(2014ZX08010002-003-002);山东省自然科学基金项目(ZR2013CM025)

摘　　要：为了解大豆ClassⅠ几丁酶基因(Chitinase gene)对不同胁迫响应的分子机制。利用PCR技术克隆了大豆ClassⅠChitinase基因的启动子片段(Gm CHI1p),序列分析表明,扩增片段(1 641 bp)与Gen Bank中的已知序列同源性达99.8%,且含有多个胁迫响应调控元件。利用GUS基因上游无启动子的表达载体p CAMBIA1391Z,构建GmCHI1p与GUS基因融合的植物表达载体pCAM-Gm CHI1p,并通过农杆菌介导法导入烟草中。在转基因烟草愈伤组织中检测到GUS活性,表明该启动子具有启动活性。对转基因烟草中的GUS活性进行初步定性分析,结果表明,GmCHI1p可驱动GUS基因在转基因烟草的根部特异性表达,而且在伤害处理的叶片中检测到GUS的强烈表达,表现出明显的根组织特异性及伤害诱导性。这种伤害诱导仅在伤害组织部位及其附近高效表达而没有被长距离传递,预计该启动子在转基因抗虫分子育种中具有巨大的应用前景。To investigate the molecular mechanism of the Class Ⅰ Chitinase gene response to virous stresses,the Class I Chitinase promoter was isolated from soybean by PCR and its sequencing analysis indicated that the amplified fragment（ 1 641 bp） was 99. 8% homologous to the ones reported in the Gen Bank database. The results showed that it included several conserved stress-responsive elements. By using the expression vector p CAMBIA1391 Z without promoter upstream of GUS gene,GmCHI1p was fused in the frame with the GUS reporter gene and the resulting construct p CAM-GmCHI1p was introduced into tobacco by Agrobacterium-mediated method. GUS activity can be detected in the Gm CHI1p-transfomants but not untransformed calli（ control）. The determinations of the GUS activities in the transgenic tobacco plants indicated that GmCHI1p could drive the GUS reporter gene to express in the roots specifically and could be significantly induced by wounding. The GmCHI1promoter was root tissue-specific and wounding-inducible in its expression. The wounding-inducible expression was occurred only in the wounding tissue or at round it but not transmitted for a long distance. It is expected to have great application prospects in the molecular breeding of transgenic insect resistance.

关 键 词：几丁质酶 启动子 根特异性 伤害诱导性 

分 类 号：Q78[生物学—分子生物学] S565.03[农业科学—作物学]