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.

1 . A method of producing a protein comprising introducing into an avian genome an exogenous nucleotide sequence comprising a fragment of the nucleotide sequence of SEQ ID NO: 67 operably linked to an exogenous coding sequence wherein the avian produces an exogenous protein. 
     
     
         2 . The method of  claim 1  wherein the protein is deposited in egg white produced by the avian. 
     
     
         3 . The method of  claim 1  comprising isolating the exogenous protein. 
     
     
         4 . The method of  claim 1  wherein the exogenous protein is secreted from an oviduct cell. 
     
     
         5 . The method of  claim 4  comprising isolating the exogenous protein. 
     
     
         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 nucleotide sequence comprises a vector. 
     
     
         8 . The method of  claim 7  wherein the vector is selected from the group consisting of a plasmid, a viral vector and an artificial chromosome. 
     
     
         9 . The method of  claim 1  wherein the exogenous protein is an antibody or an enzyme. 
     
     
         10 . The method of  claim 1  comprising isolating the exogenous protein wherein the exogenous protein is selected from the group consisting of fusion proteins, growth hormones, cytokines, structural proteins, enzymes, human growth hormone, interferon, lysozyme, and β-casein, albumin, α-1 antitrypsin, antithrombin III, collagen, factor VIII, factor IX, factor X, fibrinogen, insulin, lactoferrin, protein C, erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), tissue-type plasminogen activator (tPA), somatotropin, chymotrypsin, immunoglobulins, antibodies, immunotoxins, b-domain deleted factor VIII, factor VIIa, anticoagulants; hirudin, alteplase, tpa, reteplase, tpa, tpa—3 of 5 domains deleted, insulin, insulin lispro, insulin aspart, insulin glargine, long-acting insulin analogs, glucagons, follitropin-beta, pth, fsh, pdgh, ifn alpha2, ifn alpha2a, ifn alpha2b, inf-apha, inf-beta, inf-beta 1b, ifn-beta 1a, ifn-gamma1b, il-2, il-11, hbsag, ospa, mab directed against t-lymphocyte antigen, mab directed against tag-72, tumor-associated glycoprotein, chimeric mab directed against platelet surface receptor gpII(b)/III(a), mab directed against tumor-associated antigen ca125, mab directed against human carcinoembryonic antigen, cea, mab directed against human cardiac myosin, mab directed against tumor surface antigen psma, mab directed against hmw-maa, mab directed against carcinoma-associated antigen, mab directed against nca 90, mab directed against cd20 antigen found on surface of b lymphocytes, mab directed against the alpha chain of the il2 receptor, mab directed against the alpha chain of the il2 receptor, mab directed against tnf-alpha, mab directed against an epitope on the surface of respiratory synctial virus, mab directed against her 2, human epidermal growth factor receptor 2, human mab directed against cytokeratin tumor-associated antigen anti-ctla4, mab directed against cd 20 surface antigen of b lymphocytes dornase-alpha dnase, beta glucocerebrosidase, tnf-alpha, il-2-diptheria toxin fusion protein, tnfr-lgg fusion protein laronidase, dnaases, alefacept, tositumomab, mab, alemtuzumab, rasburicase, agalsidase beta, teriparatide, parathyroid hormone derivatives, adalimumab, anakinra, biological modifier, nesiritide, human b-type natriuretic peptide (hbnp), colony stimulating factors, pegvisomant, human growth hormone receptor antagonist, protein c, omalizumab, immunoglobulin e (lge) blocker, lbritumomab tiuxetan, ACTH, glucagon, somatostatin, somatotropin, thymosin, parathyroid hormone, pigmentary hormones, somatomedin, luteinizing hormone, chorionic gonadotropin, hypothalmic releasing factors, antidiuretic hormones, prolactin and thyroid stimulating hormone. 
     
     
         11 . A method comprising producing a protein comprising introducing a nucleotide sequence at least 95% identical to nucleotides 7665 to 11863 of SEQ ID NO: 67 operably linked to an exogenous protein coding sequence into an oviduct cell wherein the exogenous protein is expressed in the oviduct cell. 
     
     
         12 . The method of  claim 11  wherein the exogenous protein is secreted from the oviduct cell. 
     
     
         13 . The method of  claim 11  wherein the avian oviduct cell is a cell of a chicken. 
     
     
         14 . The method of  claim 11  wherein the oviduct cell is a tubular gland cell. 
     
     
         15 . The method of  claim 11  wherein the avian lysozyme gene expression controlling region comprises a nucleotide sequence at least 99% identical to nucleotides 7665 to 11863 of SEQ ID NO: 67. 
     
     
         16 . The method of  claim 11  wherein the avian lysozyme gene expression controlling region comprises a nucleotide sequence identical to nucleotides 5381 to 11863 of SEQ ID NO: 67. 
     
     
         17 . The method of  claim 11  comprising isolating the exogenous protein. 
     
     
         18 . The method of  claim 11  wherein the exogenous protein is a therapeutic protein and the therapeutic protein is isolated. 
     
     
         19 . A method of producing a therapeutic protein comprising introducing into an avian genome an exogenous nucleotide sequence comprising a fragment of the nucleotide sequence of SEQ ID NO: 67 wherein the avian produces a therapeutic protein in an oviduct cell and secretes the therapeutic protein from the oviduct cell. 
     
     
         20 . The method of  claim 20  comprising isolating the therapeutic protein.