Method for identifying genes encoding signal sequences

Abstract

The invention involves a method of identifying nucleic acid sequences encoding signal peptide-containing proteins. The method features chimeric constructs containing a KRE9 gene that lacks a signal sequence. Yeast containing chimeric KRE9 plasmid constructs that encode signal sequences are selected based on their ability to grow on media in which sucrose is the sole carbon source.

1 - 17 . (canceled)  
     
     
         18 . An isolated polypeptide selected from the group consisting of: 
 i) a polypeptide encoded by a nucleic acid molecule which hybridizes to the complement of the nucleic acid molecule consisting of the nucleotide sequence of SEQ ID NO:1 or 3 under conditions comprising washing in 0.2×SSC, 0.1% SDS at 42° C.;    ii) a polypeptide encoded by a nucleic acid molecule which hybridizes to the complement of the nucleic acid molecule consisting of the nucleotide sequence of SEQ ID NO:1 or 3 under conditions comprising hybridizing in Church buffer (7% SDS, 0.5% NaHPO 4 , 1 M EDTA, 1% BSA) at 50° C., and washing at in 2×SSC at 50° C.;    iii) a polypeptide encoded by an isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1 or 3;    iv) a polypeptide encoded by an isolated nucleic comprising an open reading frame of the nucleotide sequence of SEQ ID NO:1 or 3;    v) a polypeptide comprising the amino acid sequence of SEQ ID NO:2 or 4; and    vi) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 or 4.    
     
     
         19 . The isolated polypeptide of  claim 18 , wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to the complement of the nucleic acid molecule consisting of the nucleotide sequence of SEQ ID NO:1 or 3 under conditions comprising washing in 0.1×SSC at 68° C.  
     
     
         20 . The isolated polypeptide of  claim 18 , wherein the polypeptide is encoded by an isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1 or 3.  
     
     
         21 . The isolated polypeptide of  claim 18 , wherein the polypeptide is encoded by an isolated nucleic comprising an open reading frame of the nucleotide sequence of SEQ ID NO:1 or 3.  
     
     
         22 . The isolated polypeptide of  claim 18 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2 or 4.  
     
     
         23 . The isolated polypeptide of  claim 18 , wherein the polypeptide consists of the amino acid sequence of SEQ ID NO:2 or 4.  
     
     
         24 . The polypeptide of  claim 18 , further comprising a heterologous polypeptide.