James Eddie, Lee James M
School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA 99164-2710, USA.
Plant Cell Rep. 2006 Jul;25(7):723-7. doi: 10.1007/s00299-005-0096-z. Epub 2006 Feb 25.
This paper addresses the problem of decreasing protein expression levels in genetically modified plant cells. A modeling approach was used to explain the loss of productivity over successive generations. Using this model, productivity losses were simulated for two cell lines. Although the lines were relatively stable, the protein production level could decrease by more than 80% after a large number of generations. Motivated by this problem, a dispersion method was developed to isolate productive cells from existing cell suspensions. Dilution of transgenic cells in a feeder layer of nonproducing cells at a ratio of 1:1000 facilitated the recovery of distinct, separate daughter colonies. Applying this method, it was possible to recover high producing cell lines.
本文探讨了转基因植物细胞中蛋白质表达水平下降的问题。采用建模方法来解释连续几代生产力的损失。利用该模型，对两个细胞系的生产力损失进行了模拟。尽管这些细胞系相对稳定，但经过大量传代后，蛋白质产量水平可能会下降80%以上。受此问题的启发，开发了一种分散方法，从现有的细胞悬液中分离出有生产力的细胞。将转基因细胞以1:1000的比例稀释到非生产性细胞的饲养层中，有助于恢复明显、分离的子代菌落。应用该方法，可以获得高产细胞系。
