The effect of immobilization on cell physiology and how this determines cell metabolic performance is an important concern for developing bioprocess. This is particularly true for genetically modified microorganisms and their genetic stability. For this reason the stability and physiological state of plasmid-bearing E. coli cells were ascertained by flow cytometry. Differences in the cellular DNA and protein content (15-20%) permit discrimination of control and plasmid-bearing cells, as well as adaptation to continuous cultivation conditions in both freely suspended and immobilized states to be monitored. Moreover, the observed metabolic burden due to maintenance and over-expression of plasmid-coded genetic material and slow cell growth in poorly-viable immobilized cells were found to be the main factors contributing to strain stabilization.

译文

:固定化对细胞生理的影响以及这如何决定细胞代谢性能是发展生物过程的重要考虑因素。对于转基因微生物及其遗传稳定性尤其如此。因此,通过流式细胞术确定了带有质粒的大肠杆菌细胞的稳定性和生理状态。细胞DNA和蛋白质含量的差异(15-20%)可以区分对照细胞和带有质粒的细胞,并可以监测自由悬浮和固定状态下连续培养条件的适应性。此外,已发现由于质粒编码的遗传物质的维持和过表达以及在存活率低的固定化细胞中缓慢的细胞生长而导致的观察到的代谢负担是促成菌株稳定的主要因素。

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