Tn(pb-EGFP/Tdrd6)#Jess

A loxP site flanked neomycin resistance gene cassette was inserted into an artificial intron (flanked by splice donor and acceptor sites) inserted into the EGFP (enhanced green fluorescent protein) gene. This construct, followed by PreScission cleavage site, S-peptide and TEV cleavage site sequences, was then inserted upstream of the ATG translation start site of the mouse gene in BAC clone RP23-283P8. A cassette carrying the piggyBac inverted repeats was inserted into the vector. To integrate the transgenic construct into the genome, the vector was injected into the pronucleus of fertilized C57B6L/6 eggs together with piggyBac transposase mRNA. This creates an allele for a Localization and Affinity Purification (LAP)-tagged protein, with the N-terminally attached EGFP serving as a tag both in localization studies and in immunoprecipitation experiments. Immunoblots showed protein expression from this allele in testes. Subsequent crossing with mice carying the Tdrd6tm1Jess allele showed that the transgene was able to rescue that null allele. A number of founders were established but no line information is specified (indicated by the pound # sign). (J:242066)