Molecular Image Detail

Image

Caption

Generation of the Gnl3^tm2.1Rylt (NS^flox) and Gnl3^tm2.2Rylt (NS^null) allele. (A) The gene-targeting vector contains two loxP sites (black arrowheads) flanking the first two exons, two Frt sites (gray arrowheads) flanking a neomycin-resistance cassette (pgk-neo), and two recombineering arms. Capital letters stand for restriction sites: B, BamHI; P, PstI; H, Hind-3; RV, EcoRV; RI, EcoRI. (B) The correctly targeted ES clones yield a 14.3-kb BamHI-digested fragment from the endogenous allele and a 7.9-kb (or 8.4-kb) fragment from the NS^flox-neo (fn) allele when hybridized with the 3' (3'-P) or 5' (5'-P) external probe. The Flp-excised NS^flox (f) allele and the germ-line Cre-excised NS^null (n) allele can be verified by BamHI-digested Southern blots hybridized with the 5' external probe. (C) Two correctly targeted ES clones (6H and 7C) were identified by Southern blot and used to establish germ-line transmission of the NS^fn allele. (D) NS^fn/+ heterozygotes were removed of the pgk-neo cassette to produce NS^f/+ mice. A PCR-based genotyping strategy was designed to detect the NS^f, NS wild-type (NS⁺), NSⁿ, and Cre alleles.

Associated
Alleles

Symbol	Name
Gnl3^tm2.1Rylt	guanine nucleotide binding protein nucleolar 3; targeted mutation 2.1, Robert Y L Tsai
Gnl3^tm2.2Rylt	guanine nucleotide binding protein nucleolar 3; targeted mutation 2.2, Robert Y L Tsai

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