DNA vector with isolated cDNA gene encoding metallopanstimulin

Abstract

A novel DNA sequence is disclosed which encodes a protein associated with many human cancers. This protein is designated as metallopanstimulin-1 (MPS-1) since (1) it is associated with metal ions, particularly zinc; (2) it has been detected in numerous different types of cells; (3) it is associated with rapid cell proliferation. The MPS-1 mRNA and its encoded protein are expressed in normal cells to a much lesser degree than in premalignant or malignant tumor cells, and they are present at very low levels in senescent cells compared to young healthy cells. The DNA sequence and the protein can be used in diagnostic methods such as detection of malignant cells associated with several types of tumors. Thus, this invention discloses a method for determining the presence of certain types of malignant conditions in patients. The MPS-1 cDNA sequence has been inserted into convenient vectors, and a culture of E. coli cells containing the sequence has been deposited with the American Type Culture Collection (ATCC), under accession number ATCC 68656.

I claim: 
     
       1. An isolated nucleic acid molecule which encodes a protein, designated metallopanstimulin-1, which has the amino acid sequence shown in FIG. 4 (SEQ ID NO:2). 
     
     
       2. A genetic vector which replicates in E. coli comprising the nucleic acid molecule of claim 1. 
     
     
       3. A genetic vector capable of transfecting insect cells comprising the nucleic acid molecule of claim 1. 
     
     
       4. Transformed bacterial cells comprising the vector of claim 2. 
     
     
       5. Transfected insect cells comprising the vector of claim 3. 
     
     
       6. An isolated nucleic acid probe consisting of nucleotides 21-41 of SEQ ID NO:1 or nucleotides complementary to nucleotides 236-256 of SEQ ID NO:1. 
     
     
       7. The isolated nucleic acid probe of claim 6 which is conjugated to at least one biotin moiety. 
     
     
       8. The isolated nucleic acid probe of claim 6 which is conjugated to at least one fluorescent moiety. 
     
     
       9. The isolated nucleic acid probe of claim 6 which is radioactively labeled. .Iadd. 
     
     
       10.  An isolated nucleic acid molecule which encodes a mutein of metallopanstimulin-1 having the amino acid sequence shown in FIG. 4 (SEQ ID NO:2), wherein one or both cysteine residues at positions 64 and 77 have been replaced by another amino acid. .Iaddend..Iadd. 
     
     
       11.  A recombinant expression vector comprising the nucleic acid molecule of claim 1 or claim 10. .Iaddend..Iadd.12. A host cell transformed or transfected with an expression vector of claim 11. .Iaddend..Iadd.13. A process for preparing metallopanstimulin-1 polypeptide comprising culturing a host cell according to claim 12 under conditions promoting expression, and recovering metallopanstimulin-1 polypeptide from the culture. .Iaddend..Iadd.14. The vector of claim 11 comprising a chimeric gene wherein the nucleic acid which encodes metallopanstimulin-1 or mutein thereof is operatively linked to a nucleic acid which encodes a heterologous polypeptide. .Iaddend..Iadd.15. A host cell transformed or transfected with an expression vector of claim 14. .Iaddend..Iadd.16. A process for preparing a metallopanstimulin-1 polypeptide or mutein thereof comprising culturing a host cell according to claim 15 under conditions promoting expression, and recovering the metallopanstimulin-1 polypeptide 
     
     
        or mutein thereof from the culture. .Iaddend..Iadd.17.  A probe comprising nucleotides 21-256 of SEQ ID NO:1. .Iaddend..Iadd.18. The probe of claim 17 which is conjugated to at least one biotin unit. .Iaddend..Iadd.19. The probe of claim 17 which in conjugated to at least one one fluorescent unit. .Iaddend..Iadd.20. The probe of claim 17 which is radioactively labeled. .Iaddend.