Functional Analysis of Basic Transcription Element Binding Protein by Gene Targeting Technology

Abstract

Basic transcription element binding protein (BTEB) is a transcription factor with a characteristic zinc finger motif and is most remarkably enhanced by thyroid hormone T 3 treatment (R. J. Denver et al., J. Biol. Chem. 272:8179-8188, 1997). To investigate the function of BTEB per se and to touch on the effects of T 3 (3,5,3-triiodothyronine) on mouse development, we generated BTEB-deficient mice by gene knockout technology. Homologous BTEB؊/؊ mutant mice were bred according to apparently normal Mendelian genetics, matured normally, and were fertile. Mutant mice could survive for at least 2 years without evident pathological defects. From the expression of lacZ, which was inserted into the reading frame of the BTEB gene, BTEB showed a characteristic tissue-specific expression profile during the developmental process of brain and bone. Dramatically increased expression of BTEB was observed in Purkinje cells of the cerebellum and pyramidal cell layers of the hippocampus at P7 when synapses start to form in the brain. Although general behavioral activities such as locomotion, rearing, and speed of movement were not so much affected in the BTEB ؊/؊ mutant mice, they showed clearly reduced activity levels in rotorod and contextual fear-conditioning tests; this finding was probably due to defective functions of the cerebellum, hippocampus, and amygdala.

Subfertility, Uterine Hypoplasia, and Partial Progesterone Resistance in Mice Lacking the Krüppel-like Factor 9/Basic Transcription Element-binding Protein-1 (Bteb1) Gene

“…Klf9/Bteb1 mutant mice (Bteb1 lacZ ) were generated by insertion of the bacterial ␤-galactosidase (LacZ) gene in-frame into the first exon of the mouse Bteb1 gene as described previously (23). Mice were genotyped by PCR of genomic DNA prepared from mouse tails by overnight digestion at 55°C in Tris-HCl buffer containing Triton X-100 (0.5%) and proteinase K (50 g/ l).…”

“…Because currently available anti-BTEB1 antibodies, including ours, have so far given high nonspecific staining in immunohistochemical studies of mouse tissues (data not shown), we took advantage of the functional ␤-galactosidase gene that was inserted into exon 1 of the Bteb1 gene to determine endogenous sites of Bteb1 expression in the early pregnant uterus. A previous study (23) demonstrated that X-gal staining of various tissues from Bteb1 ϩ/lacZ mice recapitulates the endogenous expression pattern of the Bteb1 gene. X-gal stained-cryosections of uterine tissues isolated from Bteb1 ϩ/lacZ mice at 0.5, 3.5, and 6.5 dpc revealed the distinct cell type localization of this gene (Fig.…”

“…Growth Phenotype of Bteb1-deficient Mice-Bteb1 Ϫ/Ϫ mutants exhibited no obvious external phenotype and appeared to mature normally (23). Because BTEB1 has been shown to influence proliferation of cells in vitro (36), the growth rates of WT and Bteb1 Ϫ/Ϫ offspring were evaluated.…”

“…We recently identified a novel PR-interacting protein, basic transcription element-binding protein-1 (BTEB1), whose expression has been documented in reproductive and non-reproductive tissues (22,23). BTEB1, a member of the Sp/Krü p-pel-like family (KLF) of transcription factors (24,25), preferentially binds PR-B in vitro and enhances PR-mediated transactivation in endometrial epithelial cells (26).…”

Desenvolvimento De Bioconservante Natural Para Aplicação Em Alimentos

“…Proteínas obtidas a partir de indústrias de óleos vegetais em são mais suscetíveis à contaminação por Salmonella spp. (Morita et al, 2003;European Food Safety Authority, 2006). …”