DNA encoding human endothelial cell growth factors and plasmids comprising said DNA

Abstract

The present invention is directed to DNA encoding human endothelial cell growth factors, and to plasmids comprising said DNA. In particular, the invention relates to DNA encoding a cleavable signal peptide and an endothelial cell growth factor, wherein removal of said signal peptide yields a mature form of the growth factor.

What is claimed is:  
     
       1. An isolated DNA encoding a cleavable signal peptide and an endothelial cell growth factor, wherein removal of said signal peptide yields a mature form of said endothelial cell growth factor, and said endothelial cell growth factor either has the amino acid sequence of α-endothelial cell growth factor 
       (NYKKPKLLYCSNGGHFLRILPDGTVDGTRDRSDQHI QLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQ TPNEECLFLERLEENHYNTYI S K K H A E K N W F V G L K KNGSCKRGPRTHYGQKAILFLPLPVSSD) or comprises the amino acid sequence of α-endothelial  β-βendothelial cell growth factor  
       (AEGEITTFTALTEKFNLPPGNYKKPKLLYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLYYGSQTPNEECLFLERLEENHYNTYIS KKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPL PVSSD).  
     
     
       2. An isolated DNA according to  claim 1 , wherein said endothelial cell growth factor is human α-endothelial cell growth factor having the amino acid sequence 
       N Y K K P K L L Y C S N G G H F L R I L P DGTVDGTRDRSDQHI QLQLSAESVGEVYIKSTETGQYLAMNTDGLLYGSQ TPNEECLFLERLEENHYNTYISKKHAEKNWFVGLK KNGSCKRGPRTHYGQKAILFLPLPVSSD.  
     
     
       3. An isolated DNA according to  claim 1 , wherein said endothelial cell growth factor is human β-endothelial cell growth factor having the amino acid sequence 
       A E G E I T T F T A L T E K F N L P P G N Y K K P K L LYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLLYGSQT P N E E C L F L E R L E E N H Y N T Y I S KKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPL PVSSD.  
     
     
       4. An isolated DNA according to  claim 1 , wherein said signal peptide is a heterologous signal peptide. 
     
     
       5. A DNA encoding a cleavable heterologous signal peptide and an endothelial cell growth factor, wherein removal of said signal peptide yields a mature form of said endothelial cell growth factor, and said endothelial cell growth factor either has the amino acid sequence of α-endothelial cell growth factor 
       (NYKKPKLLYCSNGGHFLRILPDGTVDGTRDRSDQHI QLQLSAEVSVGEVYIKSTETGQYLAMDTDGLLYGSQ TPNEECLFLERLEENHYNTY I S K K H A E K N W F V G L K KNGSCKRGPRTHYCQKAILFLPLPVSSD) or comprises the amino acid sequence of β-endothelial cell growth factor  
       (AEGEITTFTALTAKFNLPPGNYKKPKLLYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLLYGSQTPNEECLFLERLEENHYNTYIS KKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPL PVSSD).  
     
     
       6. A DNA according to  claim 5 , wherein said endothelial cell growth factor is human α-endothelial cell growth factor having the amino acid sequence 
       N Y K K P K L L Y C S N G G H F L R I L P DGTVDGTRDRSDQHI QLQLSAESVGEVYIKSTETGQYLAMDTFDGLLYGSQ TPNEECLFLERLEENHYNTYISKKHAEKNWFVGLK KNGSCKRGPRTHYGQKAILFLPLPVSSD.  
     
     
       7. A DNA according to  claim 5 , wherein said endothelial cell growth factor is human β-endothelial cell growth factor having the amino acid sequence 
       AEGEITTFTALTEKFNLPPGNYKKPKLLYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLLYGSQT P N E E C L F L E R L E E N H Y N T Y I S KKHAEKNWFVGLKKNGSCKRGPRRTHYGQKAILFLPL PVSSD.  
     
     
       8. A plasmid comprising a DNA encoding a cleavable signal peptide and an endothelial cell growth factor, wherein removal of said signal peptide yields a mature form of said endothelial cell growth factor, and said endothelial cell growth factor either has the amino acid sequence of α-endothelial cell growth factor 
       (NYKKPKLLYCSNGGHFLRILPDGTVDGTRDRSDQHI QLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQ TPNEECLFLERLEENHYNTY I S K K H A E K N W F V G L K KNGSCKRGPRTHYGQKAILFLPLPVSSD)  
       or comprises the amino acid sequence of β-endothelial cell growth factor 
       (AEGEITTFTALTEKFNLPPGNYKKPKLLYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLLYGSQTPNEECLFLERLEENHYNTYIS KKHAQEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPL PVSSD).  
     
     
       9. A plasmid according to  claim 8 , wherein said endothelial cell growth factor is human α-endothelial cell growth factor having the amino acid sequence 
       N Y K K P K L L Y C S N G G H F L R I L P DGTVDGTRDRSDQHI QLQLSAESVGEVYIKSTETGQYLAMDTDGLYYGSQ TPNEECLFLERLEENHYNTYISKKHAEKNWFVGLK KNGSCKRGPRTHYGQKAILFLPLPVSSD.  
     
     
       10. A plasmid according to  claim 8 , wherein said endothelial cell growth factor is human β-endothelial cell growth factor having the amino acid sequence 
       A E G E I T T F T A L T E K F N L P P G N Y K K P K LLLYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLLYGSQT P N E E C L F L E R L E E N H Y N T Y I S KKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPL PVSSD.  
     
     
       11. A plasmid according to  claim 8 , wherein said signal peptide is a heterologous signal peptide. 
     
     
       12. A plasmid comprising a DNA encoding a cleavable heterologous signal peptide and human α-endothelial cell growth factor comprising  having the amino acid sequence 
       N Y K K K L L Y C S N G G H F L R I L P DGTVDGTRDRSDQHI QLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQ TPNEECLFLERLEENHYNTYISKKHAEKNWFVGLK KNGSCKRGRRTHYGQKAILFLPLPVSSD,  
       wherein removal of said signal peptide yields a mature form of said human α-endothelial cell growth factor. 
     
     
       13. A plasmid comprising a DNA encoding a cleavable heterologous signal peptide and human β-endothelial cell growth factor comprising the amino acid sequence 
       AEGEITTFTALTEKFNLPPGNYKKPKLLYCSNGGHFLR ILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETG QYLAMDTDGLLYGSQTPNEECLFLERLEENHYNTYIS KKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPL PVSSD,  
       wherein removal of said signal peptide yields a mature form of said human β-endothelial cell growth factor. 
     
     
       14. A process for expressing an endothelial cell growth factor in a host cell, comprising introducing the plasmid according to  claim 8  into the host cell. 
     
     
       15. A process for expressing a human α-endothelial cell growth factor in a host cell, comprising introducing the plasmid according to  claim 9  into the host cell. 
     
     
       16. A process for expressing a human β-endothelial cell growth factor in a host cell, comprising introducing the plasmid according to  claim 10  into the host cell. 
     
     
       17. A process for expressing an endothelial cell growth factor in a host cell, comprising introducing the plasmid according to  claim 11  into the host cell. 
     
     
       18. A process for expressing a human α-endothelial cell growth factor in a host cell, comprising introducing the plasmid according to  claim 12  into the host cell. 
     
     
       19. A process for expressing a human β-endothelial cell growth factor in a host cell, comprising introducing the plasmid according to  claim 13  into the host cell. 
     
     
       20. The process according to  claim 14 , wherein the host cell is a prokaryotic cell. 
     
     
       21. The process according to  claim 20 , wherein the prokaryotic cell is E. coli. 
     
     
       22. A process for preparing an endothelial cell growth factor, comprising transforming a host cell with a plasmid according to  claim 8 , culturing the host cell under conditions permitting expression of the endothelial cell growth factor, and recovering the endothelial cell growth factor. 
     
     
       23. A process for preparing a human α-endothelial cell growth factor, comprising transforming a host cell with a plasmid according to  claim 9 , culturing the host cell under conditions permitting expression of the human α-endothelial cell growth factor, and recovering the human α-endothelial cell growth factor. 
     
     
       24. A process for preparing a human β-endothelial cell growth factor, comprising transforming a host cell with a plasmid according to  claim 10 , culturing the host cell under conditions permitting expression of the human β-endothelial cell growth factor, and recovering the human β-endothelial cell growth factor. 
     
     
       25. A process for preparing an endothelial cell growth factor, comprising transforming a host cell with a plasmid according to  claim 11 , culturing the host cell under conditions permitting expression of the endothelial cell growth factor, and recovering the endothelial cell growth factor. 
     
     
       26. A process for preparing a human α-endothelial cell growth factor, comprising transforming a host cell with a plasmid according to  claim 12 , culturing the host cell under conditions permitting expression of the human α-endothelial cell growth factor, and recovering the human α-endothelial cell growth factor. 
     
     
       27. A process for preparing a human β-endothelial cell growth factor, comprising transforming a host cell with a plasmid according to  claim 13 , culturing the host cell under conditions permitting expression of the human β-endothelial cell growth factor, and recovering the human β-endothelial cell growth factor. 
     
     
       28. The process according to  claim 23 , wherein the host cell is a prokaryotic cell. 
     
     
       29. The process according to  claim 28 , wherein the prokaryotic cell is E. coli.