Biolistic transformation of prokaryotes: factors that affect biolistic transformation of very small cells.
Biolistic transformation of prokaryotes: factors that affect biolistic transformation of very small cells.
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DOI：
10.1099/00221287-138-1-239
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
F. Smith;P. Harpending;J. Sanford
中科院分区：
文献类型：
--
作者：
F. Smith;P. Harpending;J. Sanford
使用粒子枪技术转化了五种细菌。不需要对细胞进行预处理。优化了基因枪转化大肠杆菌的物理条件（氦气压力、靶细胞距离和间隙距离）和生物条件（细胞生长期、螺旋霉素浓度和细胞密度），并成功转化了根癌农杆菌、梨火疫病菌、斯氏欧文氏菌和假单胞菌。你好E.每0.8微克DNA，每板10（4）个。尽管其他物种的转化率较低（每0.8微克DNA小于10（2）/板），但对于每个测试的物种，在没有优化的情况下成功转化表明原核生物的生物射弹转化的广泛用途。E.大肠杆菌已经被证明是一个有用的模型系统，以确定相对湿度，颗粒大小和颗粒涂层对生物射弹转化效率的影响。
Five bacterial species were transformed using particle gun-technology. No pretreatment of cells was necessary. Physical conditions (helium pressure, target cell distance and gap distance) and biological conditions (cell growth phase, osmoticum concentration, and cell density) were optimized for biolistic transformation of Escherichia coli and these conditions were then used to successfully transform Agrobacterium tumefaciens, Erwinia amylovora, Erwinia stewartii and Pseudomonas syringae pv. syringae. Transformation rates for E. coli were 10(4) per plate per 0.8 micrograms DNA. Although transformation rates for the other species were low (less than 10(2) per plate per 0.8 micrograms DNA), successful transformation without optimization for each species tested suggests wide utility of biolistic transformation of prokaryotes. E. coli has proven to be a useful model system to determine the effects of relative humidity, particle size and particle coating on efficiency of biolistic transformation.