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Figure 3. The pMesogenin2 5 prime regulatory region drives expression of the EGFP reporter gene in PSM. A: Schematic diagrams of reporter constructs used in transgenesis. In pMsgn2-(-1501)-EGFP, the pMesognein2 5 prime regulatory region-EGFP cassette is flanked at both ends by two I-Sce I recognition sites. In pMsgn2-(-1501)-EGFP-p3U, the pMesogenin2 3 prime UTR is inserted between EGFP and the poly(A) signal (pA). In pMsgn2-(-1501+)-EGFP, the pMesogenin1 5 prime regulatory region is inserted between pMesogenin2 5 prime regulatory region and the I-Sce I recognition site. B: X. laevis pMesogenin1 and X. tropicalis pMesgenin1 5 prime regulatory regions compared using mVISTA (http://genome.lbl.gov/vista/mvista/submit.shtml) with the X. laevis sequence as the baseline. The downward bracket indicates the region added in pMsgn2-(-1501+)-EGFP. C-E: Whole-mount in situ hybridization for pMesogenin2 or EGFP. C: The endogenous pMesogenin2 expression at stage 27. D: In transgenic embryos with pMsgn2-(-1501)-EGFP, EGFP expression was observed in PSM, somites, and the head region. E: In transgenic embryos with pMsgn2-(1501+)-EGFP, EGFP expression was observed in PSM and somites, but was reduced in the head. F-H: Transverse sections at the positions indicated in C and D. |
