通过转染和显微注射对哺乳动物细胞进行遗传转化的效率取决于所转移的基因。

The efficiency of genetic transformation of mammalian cells by transfection and microinjection depends on the transferred gene.

作者信息

Strauss M, Kiessling U, Zavision B A, Povitza O N, Tikhonenko T I, Geissler E

出版信息

Biomed Biochim Acta. 1983;42(7-8):K27-34.

Abstract

The efficiency of genetic transformation of mammalian cells was analysed with respect to the kind of the transferred gene and the selective system. Plasmids pAGO and pAG60 harboring the thymidine kinase gene of Herpes simplex virus type 1 and the bacterial neomycin resistance gene, respectively, were compared concerning their ability to transform mouse Ltk-aprt- cells. Using the calcium phosphate technique the neomycin resistance gene transformed at least ten times more efficiently than the thymidine kinase gene (3 X 10(-3) versus 2 X 10(-4] whereas the difference is even more impressive following microinjection of the plasmids into the nuclei (2 X 10(-1) versus 2.5 X 10(-3]. The neomycin system also proved to be more effective in secondary gene transfer experiments and, thus, seems to be the most convenient marker for cotransfer experiments.

摘要

针对所转移基因的种类和选择系统，分析了哺乳动物细胞的基因转化效率。分别携带单纯疱疹病毒1型胸苷激酶基因和细菌新霉素抗性基因的质粒pAGO和pAG60，就其转化小鼠Ltk-aprt-细胞的能力进行了比较。使用磷酸钙技术，新霉素抗性基因的转化效率至少比胸苷激酶基因高十倍（3×10⁻³对2×10⁻⁴），而将质粒显微注射到细胞核后，差异更为显著（2×10⁻¹对2.5×10⁻³）。新霉素系统在二次基因转移实验中也被证明更有效，因此，似乎是共转移实验中最方便的标记物。