隐花色素CRY2基因片段克隆及RNAi植物表达载体的构建  

cDNA Cloning and Construction of RNAi Plant Expression Vector of the Cryptochrome Gene CRY2

作　　者：周波[1] 孙梅[1] 阎淑滑[1] 李玉花[1] 

机构地区：[1]东北林业大学花卉生物工程研究所,哈尔滨150040

出　　处：《分子植物育种》2008年第6期1182-1186,共5页Molecular Plant Breeding

基　　金：国家自然科学基金项目(30571510;30730078)资助

摘　　要：隐花色素是植物向光反应的主要的光受体,介导蓝光/近紫外光调控的反应。在拟南芥中CRY2调节去黄化反应并控制开化时间。我们根据在GenBank中已经发表的拟南芥(NM_100320)、中国大白菜(AY176689)、油菜(AJ889779)CRY2基因的cDNA保守序列设计并合成同源引物。通过RT-PCR方法从"津田"芜菁(Brassica rapa L.subsp.rapa"Tsuda")总RNA中扩增出CRY2基因的cDNA部分片段,采用gateway技术中的BP反应将其克隆到pDONR221中。测序表明,该片段与拟南芥具有87%的同源性,与油菜和中国大白菜的同源性达97%以上。采用gateway技术中的LR反应,以植物表达载体pH7GWIWG2(I)为基础,构建了由组成型启动子35S调控的CRY2基因的ihpRNAi植物表达载体。Cryptochrome is a group of flavin-type blue/ultraviolet-A light sensing photoreceptors that regulate various growth and developmental responses in plants. CRY2 regulates de-etiolation and flowering time in A rabidopsis. According to the conservative cDNA sequences of CRY2 in A rabidopsis （NM_100320）, Chinese cabbage （AY176689） and Brassica napus （AJ889779）, the homology primers were designed. Furthermore, the partial cDNA of CRY2 was amplified from total RNA of Brassica rapa L. subsp, rapa ＂Tsuda＂ by RT-PCR. The obtained sequence shows 87% identity to CRY2 ofA rabidopsis and over 97% identity to that of Chinese cabbage and Brassica napus. Moreover, the special fragment was constructed into the plant expression vector pH7GWIWG2（I） by Gateway cloning technology. The expression vector was regulated by 35S promoter and expected to transcript intron splicing hairpin CRY2 RNA and interfere to the inherent CRY2 gene.

关 键 词：隐花色素CRY2基因 RNAI 植物表达载体 

分 类 号：Q943.2[生物学—植物学] Q78