Production of recombinant ovine interferon tau using a Bombyx mori nuclear polyhedrosis baculovirus expression system

Autor:	Sj, Mann, Ge, Lamming, Pa, Fisher, Maeda S, Hj, Stewart, Ap, Flint, george mann
Rok vydání:	2002
Předmět:	Sheep Blotting Western Luteolysis In Vitro Techniques Pregnancy Proteins Bombyx Dinoprost Nucleopolyhedroviruses Recombinant Proteins Pregnancy Interferon Type I Chromatography Gel Animals Female Cloning Molecular Progesterone
Zdroj:	Europe PubMed Central
ISSN:	1049-5398
Popis:	Ovine interferon tau (oIFN-tau) is an embryonic protein of critical importance in the establishment of pregnancy in the sheep. We have produced recombinant (r) oIFN-tau using a baculovirus expression system and demonstrated the biological activity of the protein produced. Bombyx mori larvae were infected with B. mori nuclear polyhedrosis virus (BmNPV), modified by inserting a cDNA coding for oIFN-tau downstream of the strong polyhedron promoter. Following infection, antiviral activity of the haemolymph rose to a maximum of 3.6 x 10(8) u/mL (equivalent to 3 mg roIFN-tau/mL) by day 5, when haemolymph was collected and stored frozen. Control haemolymph, collected from uninfected insects at an equivalent time, contained no antiviral activity. The roIFN-tau was partially purified by gel filtration column chromatography and the presence of roIFN-tau confirmed by western blotting. The biological activity of the partially purified roIFN-tau was tested in ewes. Treatment with roIFN-tau caused a significant delay in luteolysis confirming biological potency. The results demonstrate that this system can be successfully used to produce large quantities of roIFN-tau.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid_dedup__::46205ba4721074fc904d7a12982014c1 https://pubmed.ncbi.nlm.nih.gov/12212938 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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