转基因
生物
微量注射
遗传学
质粒
表达式向量
分子生物学
基因
转基因小鼠
内部核糖体进入位点
细胞生物学
重组DNA
信使核糖核酸
翻译(生物学)
作者
Joanna L. Jankowsky,Hilda H. Slunt,Tamara Ratovitski,Nancy A. Jenkins,Neal G. Copeland,David R. Borchelt
出处
期刊:Biomolecular Engineering
日期:2001-06-01
卷期号:17 (6): 157-165
被引量:708
标识
DOI:10.1016/s1389-0344(01)00067-3
摘要
The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.
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