Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC-IDICAN, Cardenal Herrera Oria s/n, 39011 Santander, Spain.
Plasmid. 2012 Mar;67(2):174-82. doi: 10.1016/j.plasmid.2012.01.008. Epub 2012 Jan 24.
A cytometric method for the estimation of end-point conjugation rates is developed and adapted to surface conjugation. This method improves the through-put of conjugation assays based on replica-plating and results in less noisy experimental data. Although conjugation on solid surfaces deviates from ideal conditions in which cells are continuously mixed, results show that, within the limits of high initial population densities and short mating times, end-point estimates of the conjugation rates are robust measurements. They are independent of the donor/recipient ratios and, to some extent, of the sampling time. Remixing the mating population in the course of a conjugation experiment results in a boost in the frequency of transconjugants.
建立并改进了一种用于估计末端连接速率的细胞计量学方法，并将其应用于表面连接。该方法提高了基于复制平板的连接测定的通量，并且产生了噪声更小的实验数据。尽管在固体表面上的连接与细胞连续混合的理想条件不同，但结果表明，在高初始种群密度和短交配时间的限制内，连接速率的终点估计是稳健的测量。它们与供体/受体的比例无关，并且在某种程度上与采样时间无关。在连接实验过程中重新混合交配种群会导致转导子的频率增加。
