Carlson C A, Pierson L S, Rosen J J, Ingraham J L
J Bacteriol. 1983 Jan;153(1):93-9. doi: 10.1128/jb.153.1.93-99.1983.
Cells of Pseudomonas stutzeri are naturally transformed by homologous chromosomal DNA; they do not require chemical treatment to become competent. This capacity to undergo natural transformation was found to be shared by the closely related species P. mendocina, P. alcaligenes, and P. pseudoalcaligenes, but was not detectable in strains of P. aeruginosa, P. perfectomarinus, P. putida, P. fluorescens, or P. syringae. P. stutzeri could be transformed either on plates or in liquid medium. Only double-stranded chromosomal DNA was effective; single-stranded DNA and plasmid DNA were not. DNA fragments larger than 10 kilobase pairs were more effective than smaller fragments. The transformation frequency was proportional to DNA concentration from 1 ng/ml to 1 microgram/ml; higher concentrations were saturating. The maximum frequency, about 10(-4) transformants per recipient cell, was obtained with cells from a culture in the early stationary growth phase. A variety of chromosomal mutations have been transformed, including mutations to auxotrophy and to antibiotic resistance. Other systems for genetic exchange in P. stutzeri have not yet been found; transformation offers a means for the genetic analysis of this metabolically versatile organism.
施氏假单胞菌细胞可被同源染色体DNA自然转化;它们无需化学处理就能具备感受态。研究发现,密切相关的门多萨假单胞菌、产碱假单胞菌和类产碱假单胞菌也具有这种自然转化能力,但在铜绿假单胞菌、完美海单胞菌、恶臭假单胞菌、荧光假单胞菌或丁香假单胞菌的菌株中未检测到这种能力。施氏假单胞菌可在平板上或液体培养基中被转化。只有双链染色体DNA有效;单链DNA和质粒DNA无效。大于10千碱基对的DNA片段比小片段更有效。转化频率与DNA浓度在1纳克/毫升至1微克/毫升范围内成正比;更高浓度时达到饱和。在处于早期稳定生长阶段的培养物中的细胞中获得了最高频率,约为每受体细胞10^(-4)个转化子。多种染色体突变已被转化,包括营养缺陷型突变和抗生素抗性突变。尚未发现施氏假单胞菌的其他基因交换系统;转化为对这种代谢多样的生物体进行遗传分析提供了一种手段。