Jessen et al., 1998 - Modification of bacterial artificial chromosomes through Chi-stimulated homologous recombination and its application in zebrafish transgenesis. Proceedings of the National Academy of Sciences of the United States of America   95:5121-5126 Full text @ Proc. Natl. Acad. Sci. USA

Fig. 2 Southern blot analysis of three different GATA-2 BAC clones modified by the G2TC DNA fragment. Newly introduced restriction sites (Fig. 1D) were used to digest both wild-type (lanes 1) and modified (lanes 2) BAC DNA. (A) GATA-2 BAC clone 3 was digested with the indicated enzymes and probed with a 2-kb wild-type ClaI-EcoRV genomic fragment (Fig. 1B). A 9-kb wild-type EcoRV fragment became 8.8-kb and 2-kb fragments. An 11-kb wild-type XhoI-BamHI fragment became 9-kb and 2-kb fragments. A 20-kb wild-type SalI fragment became 12-kb and 8.6-kb fragments. A 10.5-kb wild-type ClaI fragment became 9.5-kb and 1.7-kb fragments. A 13.5-kb wild-type HindIII fragment became 12-kb and 2.5-kb fragments. GATA-2 BAC clones 1 and 4 (B) also contained the desired gene replacement.

Fig. 3 GFP expression from modified GATA-2 BACs in living zebrafish embryos. (A) GFP expression driven by modified BAC clone 4 in the ventral ectoderm and mesoderm of a 6-h shield stage embryo. Arrow indicates the dorsal shield. (B) GFP expression driven by modified BAC clone 4 in the posterior intermediate cell mass of a 20-h embryo. (C) GATA-2 expression pattern in an 18-h embryo as detected by RNA in situ hybridization. (D) GFP expression driven by modified BAC clone 3 in circulating hematopoietic cells of a 48-h embryo. GFP expression driven by modified BAC clone 4 in the EVL (E and G) and in a motoneuron (F) of 48-h embryos.

Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA