DSMZ-German Collection for Microorganisms and Cell Cultures, Inhoffenstr. 7b, 38124 Braunschweig, Germany.
Int J Mol Sci. 2013 Jul 22;14(7):15141-66. doi: 10.3390/ijms140715141.
Biotechnological approaches using genetic modifications such as homologous gene overexpression can be used to decode gene functions under well-defined circumstances. However, only the recording of the resulting phenotypes allows inferences about the impact of the modification on the organisms' evolutionary, ecological or economic performance. We here compare a potato wild-type cell line with two genetically engineered cell cultures homologously overexpressing Pathogenesis Related Protein 10a (pr-10a). A detailed analysis of the relative gene-expression patterns of pr-10a and its regulators sebf and pti4 over time provides insights into the molecular response of heterotrophic cells to distinct osmotic and salt-stress conditions. Furthermore, this system serves as an exemplar for the tracing of respiration kinetics as a faster and more sensitive alternative to the laborious and time-consuming recording of growth curves. The utility and characteristics of the resulting data type and the requirements for its appropriate analysis are figured out. It is demonstrated how this novel type of phenotypic information together with the gene-expression-data provides valuable insights into the effect of genetic modifications on the behaviour of cells on both the molecular and the macroscopic level.
生物技术方法可利用基因修饰,如同源基因过表达,在明确的条件下对基因功能进行解码。然而,只有记录下由此产生的表型,才能推断出修饰对生物体进化、生态或经济表现的影响。在这里,我们比较了一个马铃薯野生型细胞系和两个同源过表达病程相关蛋白 10a(pr-10a)的基因工程细胞培养物。对 pr-10a 及其调控因子 sebf 和 pti4 的相对基因表达模式进行详细的时间分析,为异养细胞对不同渗透和盐胁迫条件的分子反应提供了深入的了解。此外,该系统可作为追踪呼吸动力学的范例,与繁琐且耗时的生长曲线记录相比,它是一种更快、更敏感的替代方法。确定了所得数据类型的实用性和特征,以及对其进行适当分析的要求。结果表明,这种新型表型信息与基因表达数据一起,为遗传修饰对细胞在分子和宏观水平上的行为的影响提供了有价值的见解。