Figure 1

Generation of eIF-5A2 transgenic mice. (A) Three eIF-5A2 transgenic mouse founders (mouse No.: 10, 11, and 89) and their offspring (166-169) were determined by PCR. One mouse without eIF-5A2 transgene (No. 9) was used as negative control. Mouse fibroblast cell line NIH 3T3 and eIF-5A2 transgenic construct was used as negative (-) and positive (+) controls. (B) The transgene eIF-5A2 was mapped to one mouse chromosome site in line 11 by FISH. The metaphase spread was prepared from bone marrow lymphocyte. Arrow indicates the hybridization signals of eIF-5A2. (C) Expression of eIF-5A2 in transgenic mice was confirmed by Northern blot analysis. A human eIF-5A2 cDNA probe was used and it did not detect the endogenous eIF-5A2 mRNA in both wild-type and transgenic mice. (D) Western blot showed the overexpression of human eIF-5A2 in transgenic mice using liver lysates. The lower bands (17dD) were eIF-5A2 and the upper bands were shifted eIF-5A2 bands caused by posttranslational hypusination. (E) Expression of transgene eIF-5A2 in various tissues of transgenic mouse was detected by RT-PCR using a pair of human-specific primers.