Colosimo A, Goncz K K, Holmes A R, Kunzelmann K, Novelli G, Malone R W, Bennett M J, Gruenert D C
University of Vermont, Burlington, USA.
Biotechniques. 2000 Aug;29(2):314-8, 320-2, 324 passim. doi: 10.2144/00292rv01.
The transfer of foreign genes into eukaryotic cells, in particular mammalian cells, has been essential to our understanding of the functional significance of genes and regulatory sequences as well as the development of gene therapy strategies. To this end, different mammalian expression vector systems have been designed. The choice of a particular expression system depends on the nature and purpose of the study and will involve selecting particular parameters of expression systems such as the type of promoter/enhancer sequences, the type of expression (transient versus stable) and the level of desired expression. In addition, the success of the study depends on efficient gene transfer. The purification of the expression vectors, as well as the transfer method, affects transfection efficiency. Numerous approaches have been developed to facilitate the transfer of genes into cells via physical, chemical or viral strategies. While these systems have all been effective in vitro they need to be optimized for individual cell types and, in particular, for in vivo transfection.
将外源基因导入真核细胞，尤其是哺乳动物细胞，对于我们理解基因和调控序列的功能意义以及基因治疗策略的发展至关重要。为此，人们设计了不同的哺乳动物表达载体系统。特定表达系统的选择取决于研究的性质和目的，这将涉及选择表达系统的特定参数，如启动子/增强子序列的类型、表达类型（瞬时表达与稳定表达）以及所需的表达水平。此外，研究的成功取决于高效的基因转移。表达载体的纯化以及转移方法都会影响转染效率。人们已经开发出许多方法，通过物理、化学或病毒策略来促进基因导入细胞。虽然这些系统在体外都很有效，但它们需要针对不同的细胞类型进行优化，特别是针对体内转染进行优化。
