Nucleic acid encoding a chimeric polypeptide
DNA molecules comprising a sequence encoding the signal peptide of hGH linked to a heterologous protein and related cells, methods and fusion proteins.
1. A cultured cell comprising a nucleic acid sequence that encodes the signal peptide of human growth hormone (hGH) linked to a nucleic acid encoding a polypeptide selected from the group consisting of α-1 antitrypsin, antithrombin III, apolipoprotein E, apolipoprotein A-1, Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XIII, bone growth factor-2, bone growth factor-7, calcitonin, a catalytic antibody, DNAse, erythropoietin, FSH-β, DNAase, globin, glucagon, granulocyte macrophage colony stimulating factor, an immune response modifier, an immunoglobulin, insulin, insulinotropin, an insulin-like growth factor, interferon-β, an interferon-β nerve growth factor, interleukin-1, interleukin-3, interleukin-4, interleukin-6, interleukin-11, interleukin-12, an interleukin-2 receptor, an IL-1 receptor antagonist, a low density lipoprotein receptor, macrophage-colony stimulating factor, parathyroid hormone, protein kinase C, soluble CD4, superoxide dismutase, tissue plasminogen activator, transforming growth factor β, tumor necrosis factor, thyroid stimulating hormone β, tyrosine hydroxylase, and urokinase, wherein the nucleic acid sequence is stably integrated into the cell and provides increased expression and secretion of the polypeptide from the cell as compared to expression and secretion of the polypeptide with a signal peptide that is naturally associated with the polypeptide.
2. The cell of claim 1 , wherein the nucleic acid sequence that encodes the signal peptide of hGH includes an intron.
3. The cell of claim 2 , wherein the intron comprises the first hGH intron.
4. The cell of claim 1 , wherein the cell is a human cell.
5. The cell of claim 1 , wherein the cell is a fibroblast.
6. The cell of claim 1 , wherein the cell is an epithelial cell, an endothelial cell, a lymphocyte, a bone marrow cell, a glial cell, a hepatocyte, a keratinocyte, a muscle cell, a neural cell, a Bowes melanoma cell, a Daudi cell, a HeLa cell, an HL-60 cell, an HT1080 cell, a Jurkat cell, a KB carcinoma cell, a K-562 leukemia cell, an MCF-7 breast cancer cell, a MOLT-4 cell, a Namalwa cell, a Raji cell, an RPMI 8226 cell, a U-937 cell, a WI-38VA13 subline 2R4 cell, a 2780AD ovarian carcinoma cell, a WI-38 cell, or an MRC-5 cell.
7. A method of obtaining a polypeptide, the method comprising
(a) culturing the cell of claim 1 , and
(b) isolating the polypeptide from the cell or from the culture medium of the cell.