Genomic localization of the Z/EG transgene in the mouse genome.
The Z/EG transgenic mouse line, produced by Novak et al., displays tissue-specific EGFP expression after Cre-mediated recombination. The autofluorescence of EGFP allows the visualization of cells of interest displaying Cre recombination. The initial construct was designed such that cells without Cre recombination express the beta-galactosidase marker, facilitating counterselection. We used inverse PCR to identify the site of integration of the Z/EG transgene, to improve the efficiency of homozygous Z/EG mouse production. Recombined cells produced large amounts of EGFP protein, resulting in higher levels of fluorescence and therefore greater contrast with nonrecombined cells. We mapped the transgene to the G1 region of chromosome 5. This random insertion was found to have occurred 230-bp upstream from the start codon of the Rasa4 gene. The insertion of the Z/EG transgene in the C57BL/6 genetic background had no effect on Rasa4 expression. Homozygous Z/EG mice therefore had no obvious phenotype.
Colombo
Sophie
S
Institut Curie, Centre de Recherche, Developmental Genetics of Melanocytes, 91405, Orsay, France.
Kumasaka
Mayuko
M
Lobe
Corrinne
C
Larue
Lionel
L
D000818:Animals; D002460:Cell Line; D002874:Chromosome Mapping; D033481:Chromosomes, Mammalian; D005456:Fluorescent Dyes / Q000378:metabolism; D015870:Gene Expression; D005819:Genetic Markers; D016678:Genome; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006720:Homozygote; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D019076:Transgenes / Q000235:genetics*; D001616:beta-Galactosidase / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism
D005456:Fluorescent Dyes; D005819:Genetic Markers; C504471:Rasa4 protein, mouse; C485184:enhanced green fluorescent protein; D020703:ras GTPase-Activating Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase