Sladek T L, Maniloff J
J Bacteriol. 1983 Aug;155(2):734-41. doi: 10.1128/jb.155.2.734-741.1983.
Phenol-extracted DNA from mycoplasma virus L2 was able to transfect Acholeplasma laidlawii in the presence of polyethylene glycol. Transfection was sensitive to DNase and was most efficient with 36% (wt/vol) polyethylene glycol 8000 and cells in logarithmic growth. Virus production by the transfected cells was similar to that of the cells infected by intact virus. L2 DNA transfected A. laidlawii with a single-hit dose-response curve, reaching saturation at high DNA concentrations. Optimum transfection frequencies were about 10(-7) transfectants per L2 DNA molecule and 10(-4) transfectants per CFU. When DNA was present in saturating amounts, the number of transfectants increased linearly with the number of CFU present in the transfection mixture, suggesting that DNA uptake does not occur by a mechanism involving cell fusion. The cleavage of the superhelical mycoplasma virus L2 genome with restriction endonucleases that cleave the DNA molecule once reduced the transfection frequency. Host cell modification and restriction of transfecting L2 DNA were similar to those for infecting L2 virions.
在聚乙二醇存在的情况下，从支原体病毒L2中提取的酚法DNA能够转染莱氏无胆甾原体。转染对DNA酶敏感，在使用36%（重量/体积）的聚乙二醇8000且细胞处于对数生长期时效率最高。转染细胞产生病毒的情况与完整病毒感染的细胞相似。L2 DNA以单 hit 剂量反应曲线转染莱氏无胆甾原体，在高DNA浓度时达到饱和。最佳转染频率约为每L2 DNA分子10^(-7)个转染子和每CFU 10^(-4)个转染子。当DNA以饱和量存在时，转染子数量随转染混合物中CFU数量呈线性增加，这表明DNA摄取不是通过涉及细胞融合的机制发生的。用能切割DNA分子一次的限制性内切酶切割超螺旋支原体病毒L2基因组会降低转染频率。宿主细胞对转染L2 DNA的修饰和限制与感染L2病毒粒子的情况相似。
