Shehata T E, Marr A G
J Bacteriol. 1970 Sep;103(3):789-92. doi: 10.1128/jb.103.3.789-792.1970.
Helmstetter and Cummings devised a technique of synchronization in which cells are implanted on a membrane filter and the membrane is subjected to reverse flow of liquid medium. The cells in the effluent stream have predominantly the characteristics of newborn cells. The advantage of this technique is that the population experiences a minimum of physiological stress; hence, the behavior of the synchronous culture should reflect the normal divisional cycle. The disadvantage is that strains other than Escherichia coli B/r cannot be synchronized. We have found that a modification of the method makes it possible to synchronize several strains of E. coli, including both male and female strains, as well as Salmonella typhimurium LT2. The principal difference in technique is a prolonged period (>400 doublings) of cultivation in glucose minimal medium at 30 C and at low density (<5 x 10(6) cells/ml) prior to implantation. This precaution was taken to insure that the bacterial growth population is in a steady state of balanced growth. From the resulting synchronous growth, the distribution of interdivision times has been computed; these distributions have coefficients of variation in the range 0.18 to 0.22 and are not appreciably skewed.
赫尔姆斯泰特和卡明斯设计了一种同步技术,即将细胞接种在滤膜上,使滤膜受到液体培养基的反向流动作用。流出液中的细胞主要具有新生细胞的特征。该技术的优点是细胞群体所经历的生理应激最小;因此,同步培养物的行为应反映正常的分裂周期。缺点是除了大肠杆菌B/r菌株外,其他菌株无法实现同步。我们发现,对该方法进行改进后,可以使包括雄性和雌性菌株在内的几种大肠杆菌菌株以及鼠伤寒沙门氏菌LT2实现同步。技术上的主要差异在于,在接种前,要在30℃的葡萄糖基本培养基中以低密度(<5×10⁶个细胞/毫升)进行长时间(>400次倍增)培养。采取这一预防措施是为了确保细菌生长群体处于平衡生长的稳定状态。根据由此产生的同步生长情况,计算了分裂间隔时间的分布;这些分布的变异系数在0.18至0.22范围内,且没有明显的偏态。