Proc Natl Acad Sci U S A
December 7, 1999;
Germ-line transmission of transgenes in Xenopus laevis.
Adult Xenopus laevis frogs made transgenic by restriction enzyme-mediated integration were bred to test the feasibility of establishing lines of frogs that express transgenes. All of the 19 animals raised to sexual maturity generated progeny that expressed the transgene(s). The patterns and levels of expression of green fluorescent protein transgenes driven by a viral promoter, rat promoter, and four X. laevis promoters were all unaffected by passage through the germ line. These results demonstrate the ease of establishing transgenic lines in X. laevis.
Proc Natl Acad Sci U S A
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Figure 1 CMV:GFP transgenic tadpoles. (A) Control nontransgenic tadpole. (B–L) F1 CMV:GFP tadpoles from 10 different lines. Numbers in the right bottom corner represent the line number in Table 1. (K) Weak-expressing tadpole from line 17. (J) A strong-expressing tadpole from line 17. (M–O) Three F0 CMV:GFP tadpoles. All panels represent dorsal views of 8-day-old tadpoles taken at the same -s exposure, except for K, which was taken at ¼-s exposure. (Scale bar represents 500 μm.)
Figure 2 F1 tadpoles transgenic for tissue-specific promoters. (A) xNβT:TauGFP line 2, dorsal view of an 8-day-old tadpole. (B) xγCryst:GFP line 5, dorsal view of an 8-day-old tadpole. The arrowhead points to the fluorescent lens. The lens of the other eye was obscured because of the orientation of the tadpole. (C) xKer:GFP line 19, immunohistochemical detection of GFP (red), with 4′,6-diamidino-2-phenylindole counterstain (blue), in transverse section through the head of a stage 42 (12) tadpole. The arrowhead points to the skin and the arrow points to the brain. (D) rTα1:GFP line 13, dorsal view of an 8-day-old animal. (E) rTα1:GFP line 13, lateral view of a stage 54 tadpole. The arrowhead points to the forelimb and the arrow points to the hindlimb. [Scale bars represent 500 μm (A, B, and D), 20 μm (C), and 1 mm (E).]
Figure 3 Luciferase activity of xHSP70:GFP-luc transgenes in 18 F0 animals 8 h after heat shock (○) and 5 F1 transgenic animals (line 5) assayed at each time point after heat shock (●).
Figure 4 Comparison of 2-week-old F1 and F2 tadpoles from line 1. Two-week-old tadpoles that expressed GFP in the F1 generation (A) and F2 generation (C) were larger and showed hyperplastic brachial arches, whereas animals that did not express GFP in the F1 generation (B) and F2 generation (D) were smaller and had normal brachial arches. Each panel shows the same animal twice: in a reflected light view on top, and a GFP fluorescence view on the bottom. Brackets highlight the enlarged brachial arches. (Scale bar represents 1 mm.)
Amaya, A method for generating transgenic frog embryos. 1999, Pubmed