辣椒WRKY 转录因子基因CaRKNIF1 植物表达载体构建及番茄转化

	辣椒WRKY 转录因子基因CaRKNIF1 植物表达载体构建及番茄转化
	张春秋; 茆振川; 杨宇红; 陈国华; 谢丙炎
刊名	分子植物育种
	2010
卷号	8 期号:4 页码:713-718
关键词	WRKY 转录因子表达载体构建番茄遗传转化
ISSN号	1672-416X
其他题名	Construction of Plant Expression Vector for WRKY Transcription Factor CaRKNIF1 Gene from Pepper and its Transformation to Tomato
英文摘要	WRKY 转录因子广泛参与植物对生物和非生物的胁迫应答反应, 在植物防卫反应中起着重要作用. CaRKNIF1 是我们从辣椒HDA149 中分离得到的新WRKY 转录因子基因(GenBank 登陆号DQ180348), 本研究通过RT-PCR 扩增得到CaRKNIF1 基因的开放阅读框, 经由中间载体pEASY-T1 Simple, 将其定向连接到表达载体pCHF3 上, 得到植物表达载体pCHF3-CaRKNIF1. 采用电击法将pCHF3-CaRKNIF1 导入农杆菌菌株LBA4404, 农杆菌介导法获得了抗性番茄植株. PCR、Southern 杂交和RT-PCR 检测结果表明, CaRKNIF1 基因已经成功整合到番茄基因组中, 并正常表达. 此研究为进一步通过CaRKNIF1 基因提高转基因番茄的抗逆性奠定了基础; WRKY transcription factors are extensively involved in plants responses to biotic/abiotic stresses, playing important roles in plant defense responses. CaRKNIF1 gene (GenBank No.DQ180348) is a new WRKY transcription factor isolated from pepper HDA149. In this study, we amplified the CaRKNIF1 ORF by RT-PCR using adaptor primers and cloned it into intermediate vector pEASY-T1 Simple. Then, we cut it from the intermediate vector and linked it to expression vector pCHF3 to form plant expression vector pCHF3-CaRKNIF1 which was later transferred into Agrobacterium tumefaciens LBA4404 by electroporation. In this way, kanamycin resistant tomato lines were obtained. PCR, Southern blot and RT-PCR analysis indicated that CaRKNIF1 gene had been successfully integrated into the genome of transgenic tomato lines and was expressed normally. This work laid good foundation for enhancing tomato resistance to different stresses by expression of the CaRKNIF1 gene
学科主题	分子生物学
语种	中文
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/100656]
专题	蔬菜花卉研究所_植物保护研究室
作者单位	中国农业科学院蔬菜花卉研究所, 北京, 100081
推荐引用方式 GB/T 7714	张春秋,茆振川,杨宇红,等. 辣椒WRKY 转录因子基因CaRKNIF1 植物表达载体构建及番茄转化[J]. 分子植物育种,2010,8(4):713-718.
APA	张春秋,茆振川,杨宇红,陈国华,&谢丙炎.(2010).辣椒WRKY 转录因子基因CaRKNIF1 植物表达载体构建及番茄转化.分子植物育种,8(4),713-718.
MLA	张春秋,et al."辣椒WRKY 转录因子基因CaRKNIF1 植物表达载体构建及番茄转化".分子植物育种 8.4(2010):713-718.