摘要
采用根癌农杆菌介导法进行GFP基因的遗传转化,建立了农杆菌介导天山雪莲遗传转化体系。以天山雪莲胚性愈伤组织为受体;40 mg/L的卡那霉素为胚性愈伤组织筛选质量浓度;胚性愈伤组织预培养2 d后用含100μmol/L AS和500 mg/L脯氨酸的感染工程菌液侵染25 min,能够取得比较理想的转化效果。对8株卡那霉素抗性植株进行PCR及GFP荧光蛋白检测,共获得6株阳性植株。结果表明GFP基因已整合到天山雪莲的基因组中并得到表达。
A genetic transformation system of embryogenic callus（Saussurea involucrata） mediated by Agrobacterium tumefaciens was established for GFP（green fluorescent protein） gene.The result showed that efficient transformation was obtained with embryogenic callus as acceptor,kanamycin of 40 mg/L was used to screen embryogenic calli.Engineering bacteria witch contain 100 μmol/L AS and 500 mg/L proline infected for 25 min after embryogenic calli was pre-cultured 2 d.Eight resistant plants were to confirm by PCR and detection of GFP fluorescent protein and 6-positive plants obtained,which indicated that GFP gene had integrated into the Saussurea involucrata genome and expressed.
出处
《西北农业学报》
CAS
CSCD
北大核心
2010年第7期84-88,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
国家“863”重点项目“特殊生境植物资源的开发利用技术”(2007AA021401)
兵团重点项目“植物抗寒基因的克隆”(NKB025DXNK01SW)
关键词
天山雪莲
根癌农杆菌
遗传转化
GFP基因
Saussurea involucrate
Agrobacterium tumefaciens
Genetic transformation
GFP gene
