PMC:1253828 / 38917-40329 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/1253828","sourcedb":"PMC","sourceid":"1253828","source_url":"http://www.ncbi.nlm.nih.gov/pmc/1253828","text":"Deletion of Mtf1 in adult mouse liver. (a) Generation of Mtf1 conditional knockout mice. The targeted allele was obtained by homologous recombination of wild-type (wt) allele and targeting vector in ES cells. Removal of the neomycin cassette (NEO) by Cre recombinase led to the conditional knockout allele Mtf1loxP. Conditional Cre-mediated deletion of exons 3 and 4 (Mtf1Δ) results in loss of function via loss of an essential part of the DNA-binding domain and the generation of a new stop codon right after exon 2. Exons 3 to 7 of Mtf1 are indicated by grey boxes, loxP sites by black triangles. TK, thymidine kinase cassette. Restriction enzymes: San, SanDI; B, BbvCI; S, SrfI; H, HpaI. The HpaI site indicated by the crossed H was lost during the cloning procedure for the targeting vector. (b) RT–PCRs with total liver RNA from pI–pC-induced male Mtf1Mx-cre or Mtf1loxP mice. The used primer pair results in products of 589 bp and 218 bp with full-length mRNA and mRNA without exons 3 and 4, respectively. (c) EMSA with liver protein extract of a pI–pC-induced male Mtf1Mx-cre or Mtf1loxP mouse. MTF-1 protein–DNA complex formation was tested with 32P-labeled MRE consensus oligonucleotide MRE-s. Specificity of binding was verified with excess of unlabeled competitor MREd or unrelated Gal4 oligonucleotide; Sp1 bandshifts with 32P-labeled Sp1 consensus oligonucleotide were included as a loading control.","tracks":[]}