Production of Transgenic Avians Using Improved Retroviral Vectors

Abstract

A transgenic avian containing in its genome an exogenous nucleotide sequence which includes a promoter component and a vector with reduced promoter interference wherein the exogenous nucleotide sequence is integrated into the genome and the avian.

What is claimed is: 
     
         1 . A method of producing an exogenous protein comprising:
 producing a transgenic avian whose genome comprises a self-inactivation (SIN) vector, wherein said vector comprising:
 i) an avian leukosis virus (ALV)-based 5′ long terminal repeat (LTR); and 
 ii) a nucleic acid sequence encoding an exogenous protein operably linked to a promoter that causes expression in oviduct cells of the transgenic avian, 
   wherein said genome does not contain the nucleic acid sequence set forth from position 1 to position 173 of SEQ ID NO: 29 or a portion thereof, which inactivates transcription of a viral sequence in the vector, and wherein said transgenic avian produces the exogenous protein in egg white.   
     
     
         2 . The method of  claim 1 , wherein the said ALV-based LTR comprises the nucleotide sequence position 174 to position 346 of SEQ ID NO: 29, or a nucleic acid sequence having at least 90% identity thereto. 
     
     
         3 . The method of  claim 1 , wherein the exogenous coding sequence encodes a human protein. 
     
     
         4 . The method of  claim 1 , wherein the exogenous coding sequence encodes a therapeutic protein. 
     
     
         5 . The method of  claim 1 , wherein the promoter comprises a functional promoter sequence of a promoter selected from the group consisting of avian ovalbumin promoter, avian ovomucoid promoter, avian lysozyme promoter and avian conalbumin promoter. 
     
     
         6 . The method of  claim 1 , wherein the avian is selected from the group consisting of a chicken, a turkey and a quail. 
     
     
         7 . The method of  claim 1 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 25 or a nucleotide sequence having at least 90% identity thereto. 
     
     
         8 . The method of  claim 7 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 26 or a nucleotide sequence having at least 90% identity thereto. 
     
     
         9 . The method of  claim 1 , wherein said method further comprising isolating the exogenous protein from egg white of the avian. 
     
     
         10 . A transgenic avian whose genome comprises a self-inactivation (SIN) vector, said vector comprising:
 i) an avian leukosis virus (ALV)-based 5′ long terminal repeat (LTR) having a deletion of the nucleic acid sequence set forth from position 1 to position 173 of SEQ ID NO: 29 or a portion thereof; and   ii) a nucleic acid sequence encoding an exogenous protein operably linked to a promoter that causes expression in oviduct cells of the transgenic avian,
 wherein the deletion inactivates transcription activity of a viral sequence in the vector, and wherein said transgenic avian produces the exogenous protein in egg white. 
   
     
     
         11 . The transgenic avian of  claim 10 , wherein the promoter comprises a functional promoter sequence of a promoter selected from the group consisting of avian ovalbumin promoter, avian ovomucoid promoter and avian lysozyme promoter. 
     
     
         12 . The transgenic avian of  claim 10 , wherein the avian is selected from the group consisting of a chicken, a turkey and a quail. 
     
     
         13 . The transgenic avian of  claim 10 , wherein the said ALV-based LTR comprises the nucleotide sequence position 174 to position 346 of SEQ ID NO: 29, or a nucleic acid sequence having at least 90% identity thereto. 
     
     
         14 . The transgenic avian of  claim 10 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 25, or a nucleotide sequence having at least 90% identity thereto. 
     
     
         15 . The transgenic avian of  claim 14 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 26, or a nucleotide sequence having at least 90% identity thereto. 
     
     
         16 . The transgenic avian of  claim 10 , wherein the exogenous coding sequence encodes a human protein. 
     
     
         17 . The transgenic avian of  claim 10 , wherein the exogenous coding sequence encodes a therapeutic protein. 
     
     
         18 . An egg laid by the transgenic avian of  claim 10 .