Methods for producing a protein using an avian lysozyme promoter

Abstract

The invention includes methods of producing proteins in transgenic avians containing nucleic acids in their genome which contain an exogenous lysozyme gene expression controlling nucleotide sequence which typically is linked to a polynucleotide encoding a heterologous polypeptide.

Description

[0001]This application is a continuation of U.S. patent application Ser. No. 11 / 699,257, filed Jan. 26, 2007, which is a continuation-in-part of U.S. patent application Ser. No. 10 / 114,739, filed Apr. 1, 2002, which claims the benefit from provisional application Ser. No. 60 / 351,550 filed Jan. 25, 2002 and is a continuation-in-part of U.S. patent application Ser. No. 09 / 922,549, filed Aug. 3, 2001, which claims the benefit of provisional application Ser. No. 60 / 280,004, filed Mar. 30, 2001. The disclosure of U.S. patent application Ser. Nos. 11 / 699,257; 10 / 114,739 and 09 / 922,549 are incorporated herein in their entirety by reference.FIELD OF THE INVENTION[0002]The present invention relates generally to the use of avian lysozyme gene expression control or controlling regions, for example, from the chicken. More specifically, the invention relates to recombinant nucleic acids and expression vectors, transfected cells and transgenic animals, in particular transgenic avians such as tran...

AI Technical Summary

Benefits of technology

[0024]In one embodiment, an isolated nucleic acid of the present invention is useful for reducing the chromosomal positional effect of a transgene operably linked to the lysozyme gene expression control region and transfected into a recipient cell. By isolating a region of the avian genome extending from 5′ upstream of a 5′ MAR of the lysozyme locus to the junction between the signal peptide sequence and a polypeptide-encoding region, cis-elements are also included to allow gene expression in a tissue-specific manner. The lysozyme promoter region of the present invention, therefore, will allow expression of an operably linked heterologous nucleic acid insert in a transfected avian cell such as, for example, an oviduct cell.

[0030]The recombinant DNA of the invention may comprise the chicken lysozyme 3′ domain operably linked to the nucleic acid insert encoding a polypeptide. The 3′ domain may include a 3′ untranslated region, a polyadenylation signal and a 3′ MAR that may reduce positional variation in transgenic avians.

Problems solved by technology

These procedures have had limited success and may require lactating animals, with the attendant costs of maintaining individual animals or herds of large species, including cows, sheep, or goats.

However, although individual cis-regulatory elements have been isolated and sequenced, together with short regions flanking DNA, the entire nucleic acid sequence comprising the functional 5′ upstream region of the lysozyme gene has not been determined in its entirety and therefore not employed as a functional promoter to allow expression of a heterologous transgene.

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