Transgene Requirements

Transgenes should include:

Promoter, with or without enhancer

Gene to be expressed, which may be a reporter gene

Splice donor and acceptor sequences flanking an intron, these can be from another gene, e.g. beta-globin or SV40 t antigen

Termination/polyadenylation sequences, these can be from another gene, e.g. beta-globin or SV40 t antigen

Transgenes must be excised from the bacterial plasmid sequences in order to be expressed in mice

This is because the prokaryotic cloning vector sequences inhibit expression of eukaryotic genes introduced into the mouse genome. See Hogan et al. Manipulating the Mouse Embryo, and Townes, T.M. EMBO J. 4:1715-1723 (1985).

You must design your transgene so that the your sequences can be excised and purified away from the plasmid vector sequences. You will do the restriction digestion, check a small portion on a gel, submit the tube to us, and we will carry out the gel electrophoresis and purification.