Page W J, Sadoff H L
J Bacteriol. 1976 Mar;125(3):1088-95. doi: 10.1128/jb.125.3.1088-1095.1976.
Azotobacter vinelandii (ATCC 12837) became competent to be transformed by exogenous deoxyribonucleic acid towards the end of exponential growth. Competence in wild-type and nitrogenase auxotrophic (nif-) strains was repressed by the addition of ammonium salts or urea to the transformation medium. Transformation of wild-type cells and nif- strains was optimal on nitrogen-free or nitrogen-limiting medium, respectively. Transformation of wild-type cells also was enhanced when the transformation medium had low molydbate content. During the development of competence, nitrogen was growth limiting, whereas carbon (glucose) was in excess. Carbon source shift-down was not effective in inducing competence. Shifting glucose-grown wild-type cells to medium containing 0.2% beta-hydroxybutyrate initiated encystment and also induced competence. The addition of glucose to this medium blocked encystment and early competence induction and reduced the transformation frequency to the basal level. Cyclic adenosine 3',5'-monophosphate induced competence in wild-type nitrogen-fixing cells and increased the transformation frequency 1,000-fold over the basal level. Exogenous cyclic adenosine 3',5'-monophosphate however, did not reverse nitrogen repression of competence in ammonia-grown wild-type or nif- strains.
维涅兰德固氮菌(ATCC 12837)在指数生长后期能够被外源脱氧核糖核酸转化。在转化培养基中添加铵盐或尿素会抑制野生型和固氮酶营养缺陷型(nif-)菌株的感受态。野生型细胞和nif-菌株的转化分别在无氮或氮限制培养基上最为理想。当转化培养基中钼酸盐含量较低时,野生型细胞的转化也会增强。在感受态形成过程中,氮是生长限制因素,而碳(葡萄糖)过量。碳源下调对诱导感受态无效。将在葡萄糖上生长的野生型细胞转移到含有0.2%β-羟基丁酸的培养基中会引发包囊化并诱导感受态。向该培养基中添加葡萄糖会阻止包囊化和早期感受态诱导,并将转化频率降低到基础水平。环腺苷酸3',5'-单磷酸在野生型固氮细胞中诱导感受态,并使转化频率比基础水平提高1000倍。然而,外源环腺苷酸3',5'-单磷酸并不能逆转氨培养的野生型或nif-菌株中感受态的氮抑制。