Figure 3
From: Transposon mediated transgenesis in a marine invertebrate chordate: Ciona intestinalis
Creation of stable transgenic lines with Minos and mutant screening. Mixture of a transposon vector containing a promoter-gfp cassette and transposase mRNA is introduced into Ciona one-cell embryos by microinjection or electroporation. These animals are cultured until gamete maturation. Generally, sperm from Minos-introduced individuals are used to fertilize wild-type eggs to obtain F1 family individuals, which are screened to find green fluorescent protein (GFP)-positive animals. The GFP-positive animals are selected for further culture, thereby establishing transgenic lines. The F1 animals with matured eggs and sperm are subjected to self-fertilization to obtain F2 family individuals, which are screened to identify mutant phenotypes. ORF, open reading frame.