农杆菌介导法将pac1基因转入烟草的研究

以烟草叶片为外植体,利用农杆菌介导法首次将pac1基因转入吉林省主栽烟草品种吉烟9号、龙江911-21和云烟87中.经过卡那霉素抗性筛选,初步获得吉烟9号转化植株43株、龙江911-21转化植株61株、云烟87转化植株43株.经PCR检测,转化烟草吉烟9号的PCR阳性率为21/43,龙江911-21的PCR阳性率为32/61,云烟87的PCR阳性率为24/43.经PCR检测为阳性的植株均能扩增出与目的片断大小一致的特异性条带,初步表明pac1基因已经转到这些烟草基因组中.从PCR扩增稳定的转基因烟草中随机抽取5株,同时以非转基因烟草为对照,进行Southern Blot分析,结果表明目的基因pac1己经被成功地整合到烟草基因组中.

Transformation of pac 1 Gene into Tobacco via Agrobacterium-Mediated Method

In this paper,tobacco seedling leaves were used as explants and pac1 gene was transferred into important tobacco cultivars Longjiang 911-21,Jiyan 9 and Yunyan 87 by agrobacterium-mediated method for the first time.43 plants of Jiyan 9,61 plants of Longjiang 911-21 and 43 plants of Yunyan 87 were obtained by Kanamycin resistance selection.Of all the Kanamycin positive plants,positive ratio of Yunyan 9 was 21 to 43,that of Longjiang 911-21 was 32 to 61 and that of Yunyan 87 was 24 to 43 and specific corresponding fragment was amplified by PCR detection.It showed that pac1gene has been transferred into tobacco.Five tobaccos selected randomly from PCR positive plants were used for Southern Blot analysis with non transgenic tobacco as control.The results indicated that pac1gene has been integrated into genome DNA of tobacco successfully.