Bovine casein gene cluster belongs to the best studied regions of the bovine genome. However, molecular basis of the regulation of casein gene expression is still of great interest for the advancement of milk production. Identification of crucial regulatory regions governing casein gene expression would provide valuable information for marker assisted selection in dairy cattle. In our study we performed comparative analysis of the bovine beta-and kappa casein gene promoter sequences with the regulatory sequences from some other species. In addition, we used homologous mammary gland derived cell culture and luciferase reporter gene system to confirm the functionality of the proximal beta and kappa casein promoters. The longer kappa casein promoter (2064 bp) showed the highest expression level, followed by the short kappa casein promoter (925 bp) and beta casein promoter (1692 bp). Here we demonstrate the suitability of the bovine mammary gland derived cell line BME UV1 for transient gene expression under transcriptional control of the bovine casein gene promoters and compare functionality of different fragments of bovine beta-and kappa casein gene promoters using homologous in vitro system. Zusammenfassung Titel der Arbeit: Funktionsanalyse der Rinder β-und κ-Kaseingenpromotoren mit Hilfe der homologen Euterzelllinie Kasein Gen Cluster beim Rind gehören zu den am besten untersuchten Teilen des Rindergenoms. Trotzdem herrscht noch immer groβes Interesse an detaillierten Studien über die molekulare Basis der Regulation der Kaseingen-Expression zur Steigerung der Milchproduktion. Die Entdeckung der Hauptregulationsmechanismen der Kaseingen-Expression wäre von groβer Bedeutung für die markergestützte Selektion der Milchkühe. In unserer Studie haben wir die Promotorsequenzen der Rinder beta-und kappa-Kaseingene im Vergleich zu einigen anderen Säugetierarten untersucht. Die Rinder-Euterzellen BME UV1 und das Luziferase Reportergen System wurden für die Bestätigung der Funktionalität der Rinder β-und κ-Kaseingenpromoters in vitro eingesetzt. Der längere κ-Kaseingen Promoter (2064 bp) hat die stärkste Expression gezeigt, gefolgt von dem kurzen κ-Kaseingen Promoter (925 bp) und dem β-Kaseingen Promoter (1692 bp). Hier können wir demonstrieren, dass die Rindereuterzelllinie BME UV1 für die vorübergehende Genexpression unter der Kontrolle der Rinderkaseingenpromotoren geeignet ist. Die Funktion von verschiedenen Teilen der Rinder β− und κ-Kaseingenpromotern wurde in einem homologen in vitro System verglichen.
The toxicity of the mixtures of parathion (PA), toxaphene (TOX) and/or 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in ICR male mice (21-24 g) by oral intubation, in corn oil, daily for up to 14 days. On Day 15, the exposure was discontinued, and animals were monitored for an additional period of 7 days for the possible reversibility of the toxicity. The body weight gain decreased with the mixtures, as well as with the individual agricultural chemicals (ACs), during the 14-day period. The cholinesterase (ChE) activity in the serum and brain was inhibited in the animals of the groups of PA (1-10 mg kg-1) and PA (5 mg kg-1)-containing mixtures. TOX (50-200 mg kg-1) caused initial inhibitory effects of 20-65% on the serum ChE (Day 1) before producing increases of 53-64% in the enzyme activity by Day 15, with little effects on the brain ChE levels. 2,4-D (50-200 mg kg-1) resulted in significantly elevated levels of the serum ChE, with substantial decreased in the brain ChE activity. The serum glutamic pyruvic transaminase level was up (38-630%) in TOX (50 mg kg-1), 2,4-D (50 mg kg-1) or their mixture group. No pathological changes at the light microscopic level in the brain and liver were noticed. TOX and TOX-containing mixtures significantly increased the liver/body weight ratio and decreased the pentobarbital (60 mg kg-1, i.p., in saline)-induced sleep.(ABSTRACT TRUNCATED AT 250 WORDS)
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