DNA encoding human alpha and beta subunits of neuronal nicotinic acetylcholine receptor, cells transformed therewith, and a recombinant cell line expressing a functional human alpha7 subunit of neuronal nicotinic acetylcholine receptor

Abstract

Isolated nucleic acid molecules, i.e., DNA or RNA encoding human neuronal nicotinic acetylcholine receptor α and β subunits, mammalian and amphibian cells containing said DNA, methods for producing α and β subunits and recombinant (i.e., isolated or substantially pure) α subunits and β subunits are provided. In addition, cells expressing various multimeric combinations of subunits (i.e., α 1 , α 2 α 3 α 4 α 5 α 6 and/or α 7 in combination with at least one of an α and β subunit are also provided. A recombinant, non-human cell line expressing the human α 7 subunit of nAChR is also disclosed.

1 - 13 . (canceled)  
     
     
         14 . A stably transfected rodent cell line which has been engineered to express a heterologous protein, said cell line comprising a host cell transformed or transfected with a heterologous nucleic acid molecule comprising a sequence of nucleotides or ribonucleotides that inducibly express an α 7  subunit of a human neuronal nicotinic acetylcholine receptor, wherein said α 7  subunit comprises a sequence of nucleotides selected from the group consisting of: 
 (a) a sequence of nucleotides as ser forth in SEQ ID No: 11 which encode a human α 7  subunit.    (b) a sequence of nucleotides that encode a polypeptide as set forth in SEQ ID No: 12;    (c) a sequence of nucleotides degenerate with the human α 7  subunit polypeptide encoding sequence of (a) or (b).    
     
     
         15 . The stable transfected cell line according to  claim 14 , wherein the cell line is derived from rat pituitary tumor tissue.  
     
     
         16 . The stably transfected cell line as claimed in  claim 14  wherein the rodent cell line is a mouse GH 4 C 1  cell line.  
     
     
         17 . The cell line according to  claim 14  wherein the heterologous protein is a functional human neuronal nicotinic acetylcholine receptor.  
     
     
         18 . The cell line according to  claim 14 , further comprising a marker gene, wherein expression of the marker gene indicates expression of the heterologous protein.  
     
     
         19 . The cell line according to  claim 14 , wherein the heterologous nucleic acid molecule is contained within an expression vector.  
     
     
         20 . The stable transfected cell line according to  claim 14 , wherein the polypeptide of SEQ ID NO: 12 is the only heterologous acetylcholine receptor subunit expressed by the cell.  
     
     
         21 . A recombinant host cell comprising a heterologous nicotinic acetylcholine receptor that comprises a subunit encoded by a heterologous nucleic acid molecule comprising a sequence of nucleotides or ribonucleotides as set forth in SEQ ID No: 11.  
     
     
         22 . A recombinant host cell comprising a heterologous nicotinic acetylcholine receptor that comprises a subunit encoded by a heterologous nucleic acid molecule wherein said nucleic acid molecule encodes a polypeptide comprising the sequence of amino acids as set forth in SEQ ID No: 12.  
     
     
         23 . The host cell according to  claim 22 , wherein the polypeptide of SEQ ID NO: 12 is the only heterologous acetylcholine receptor subunit expressed by the cell.