Production of recombinant proteins in semen

• US 6,201,167 B1
• Filed: 01/27/1999
• Issued: 03/13/2001
• Est. Priority Date: 04/03/1997
• Status: Expired due to Fees

• Current Assignee: Universite Laval
• Original Assignee: Universite Laval
• Inventors: Pothier, François
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US09/237,974
• Time in Patent Office: 776 Days
• Field of Search: 800/13, 800/18, 800/4, 435/69.1
• US Class Current: 800/18
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2227/108 Swine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination

Abstract

The present invention relates to a method for the production and secretion into animal'"'"'s semen of an exogenous recombinant protein comprising the steps of: a) producing a non-human transgenic animal characterized by an expression system comprising a promoter specific for the genital tract or accessory glands operatively linked to an exogenous DNA sequence coding for the recombinant protein through a DNA sequence coding for a signal peptide effective in secreting and maturing the recombinant protein in genital tract tissue; b) collecting semen produced by the non-human transgenic animal; and c) isolating the exogenous recombinant protein from the semen.

Claims

1. A method for the production of a heterologous protein in semen of a mouse comprising the steps of:

production a transgenic mouse whose genome comprises an expression system comprising in operable association a p12 promoter, a DNA sequence encoding a signal peptide and a DNA sequence encoding a protein heterologous to the promoter, wherein expression of said DNA sequences results in the production and secretion of said protein of interest into the mouse'"'"'s semen to a detectable level; and

collecting semen produced by said transgenic mouse.

2. The method according to claim 1, wherein said expression system further comprises a 3′

untranslated regions downstream of the DNA sequence encoding said heterologous protein.

3. The method according to claim 1, wherein said expression system further comprises a 5′

untranslated regions between said promoter and said DNA sequence encoding a signal peptide.

4. The method according to claim 1, wherein said promoter is selected from the group consisting of p12, p25, kallikreins, PSA, SBP-C and secretory protein IV promoters.

5. The method according to claim 1, wherein said recombinant protein is selected from the group consisting of, catalase, mono- and bi-specific antibodies, immunoglobulins, cytokines, coagulation factors, tissue plasminogen activator, GM-CSF, erythropoietin, thrombopoietin, alpha-1 antitrypsin, animal growth hormones, cell surface proteins, insulin, interferons, lipases, antiviral protein, antibacterial protein, bacteriocins, peptide hormones, lipocortins and epidermal growth factor.

6. A transgenic mouse whose genome comprises in operable association a p12 promoter, a DNA sequence encoding a signal peptide and a DNA sequence encoding a protein heterologous to the promoter, wherein expression of said DNA sequences results in the production and secretion of said protein of interest into the mouse'"'"'s semen to a detectable level.