陆地棉超氧化物歧化酶基因的克隆与烟草转化  

Cloning of Superoxide Dismutase Gene from Upland Cotton and Expression in Tobacco

作　　者：胡根海[1] 喻树迅[2] 范术丽[2] 宋美珍[2] 马淑娟[2] 

机构地区：[1]河南科技学院,河南新乡453003 [2]中国农业科学院棉花研究所,河南安阳455004

出　　处：《西北植物学报》2007年第11期2169-2174,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家"863"计划资助项目(2002AA241021);国家"973"项目(2004CB117300)

摘　　要：以陆地棉‘中棉所36’和烟草‘NC89’为材料,采用RT-PCR方法,从陆地棉中克隆到了Cu/Zn-SOD的cD-NA编码区,经植物表达载体pBI-SOD,导入根癌农杆菌LBA4404,对烟草叶片进行遗传转化,经卡那霉素筛选,获得正确的Cu/Zn-SOD的cDNA编码区,构建了转基因载体,并成功地获得2株转基因烟草;经PCR、Southern blotting和SOD酶活性等检测,证明Cu/Zn-SOD基因已经整合到烟草的基因组并表达。本试验结果为转基因棉花的进一步研究奠定了基础。The ‘CRI36 ＇ （Gossypium hirsutum L. ） and ‘NC89＇ （Nicotiana tabacum L. ） were used as experi- mental materials. The cotton chloroplast copper/zinc superoxide dismutase （Cu/Zn-SOD） cDNA was cloned by RT-PCR from upland cotton. The plant expression vector pIAI-SOD was constructed and trans- ferred into LBA4404 （Agrobacterium turnefaciens L. ）. The tobacco leaves were transformed and selected on the culture medium with kanamycin. The results confirmed that the gene coding-region from cDNA am- plification products was obtained. The plant expression vector was constructed and 2 stains of the trans- formed plants were obtained. The results confirmed that the Cu/Zn-SOD gene had stably integrated into the tobacco genome and been expressed with PCR, Southern blotting and enzyme activity measurement. The study has laid the stabile foundation for cotton transformation.

关 键 词：棉花 超氧化物歧化酶 植物表达载体 

分 类 号：Q789[生物学—分子生物学]