Transgenic animals expressing salivary proteins

Abstract

The invention provides a transgenic animal having within its genome a transgene construct for gastrointestinal tract specific expression of a protein. In a preferred embodiment the protein is a phytase or a homologue thereof. Such proteins may be heterologous and may be specifically expressed in the salivary gland of the animal by operably linking the nucleic acid sequence encoding the protein with regulatory sequence including a salivary gland protein promoter/enhancer. Also provided are methods of expressing and producing proteins using such nucleic acid constructs. Further, antibodies specific to such proteins and immunological diagnostic kits are also provided.

1 - 35 . (canceled)  
     
     
         36 . A method for expressing a phytase in a non-human animal, said method comprising: 
 a) constructing a nucleic acid sequence including a transgene construct comprising: 
 i) a transgene encoding said phytase, and  
 ii) at least one regulatory sequence for gastrointestinal tract specific expression of said protein, and  
   b) transfecting the animal with said nucleic acid sequence;    whereby said animal carries within the genome of its somatic and germ cells said transgene construct and wherein said animal expresses said phytase in its gastrointestinal tract and wherein the animal is selected from the group consisting of pigs, goats, sheep, cows, horses, fish and poultry.    
     
     
         37 . The method of  claim 36  wherein said transgene construct results in salivary gland or pancreatic gland specific expression of said phytase.  
     
     
         38 . The method of  claim 37  wherein said regulatory sequence provides for salivary gland specific expression of said phytase.  
     
     
         39 . The method of  claim 38  wherein said salivary gland is a parotid gland, submaxillary, or a submandibular gland.  
     
     
         40 . The method of  claim 38  wherein said phytase is expressed in the saliva of said mammal.  
     
     
         41 . The method of  claim 38  wherein said transgene construct comprises a nucleic acid sequence according to SEQ ID NO:3, SEQ ID NO:5; or SEQ ID NO:7.  
     
     
         42 . The method of  claim 38  wherein said transgene construct comprises a nucleic acid sequence according to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4, or SEQ ID NO:6.  
     
     
         43 . The method of  claim 38  wherein said animal is a pig.  
     
     
         44 . A nucleic acid molecule comprising (a) a nucleic acid sequence encoding a phytase operably linked to (b) at least one regulatory sequence for gastrointestinal tract specific expression of said phytase.  
     
     
         45 . The molecule of  claim 44  wherein said at least one regulatory sequence comprises a salivary protein promoter/enhancer sequence, whereby expression of said protein is salivary gland specific.  
     
     
         46 . The molecule of  claim 45  wherein said salivary protein promoter/enhancer sequence comprises a parotid secretory protein (PSP) promoter/enhancer, a proline-rich protein (PRP) promoter/enhancer, a salivary amylase promoter/enhancer, or a SV40 promoter/enhancer.  
     
     
         47 . The molecule of  claim 44  wherein said molecule comprises a nucleic acid sequence according to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4, or SEQ ID NO:6.  
     
     
         48 . The molecule of  claim 44  wherein said molecule includes a nucleic acid sequence according to SEQ ID NO:3, SEQ ID NO:5; or SEQ ID NO:7.  
     
     
         49 . An antibody specific to a protein expressed by a nucleic acid sequence according to SEQ ID NO:3, SEQ ID NO:5; or SEQ ID NO:7.  
     
     
         50 . The antibody of  claim 49  wherein said antibody is monoclonal.  
     
     
         51 . The antibody of  claim 49  wherein said antibody is polyclonal.  
     
     
         52 . A hybridoma secreting the antibody of  claim 50 .  
     
     
         53 . A host cell transfected with molecule according to any one of  claim 44 .  
     
     
         54 . The host cell of  claim 53  wherein said cell is an animal cell.  
     
     
         55 . A diagnostic kit for immunologically detecting a protein expressed by a nucleic acid sequence according to SEQ ID NO:3, SEQ ID NO:5; or SEQ ID NO:7, the kit including an antibody specific to said protein.  
     
     
         56 . The kit of  claim 55  wherein said antibody is monoclonal.  
     
     
         57 . The kit of  claim 56  wherein said antibody is polyclonal.  
     
     
         58 . A kit for detecting phytase in a sample from a non-human animal prepared according to  claim 36 , said kit comprising an antibody that binds to phytase.  
     
     
         59 . The kit according to  claim 58  wherein the antibody binds to a protein expressed by a nucleic acid sequence according to SEQ ID NO: 3, SEQ ID NO: 5; or SEQ ID NO: 7.